Large-scale methods of uniformly coating packaging surfaces with a volatile antimicrobial to preserve food freshness

ABSTRACT

The present application relates to large-scale methods of uniformly coating packaging surfaces with a benzoxaborole compound.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. patent application Ser. No.16/707,516, filed Dec. 9, 2019, which is a continuation-in-part of U.S.patent application Ser. No. 15/485,500, filed Apr. 12, 2017, whichclaims the benefit of priority under 35 U.S.C. § 119(e) of U.S.Provisional Patent Application No. 62/323,247, filed Apr. 15, 2016, thecontent of which is incorporated by reference in its entirety.

This application is a continuation of U.S. patent application Ser. No.16/707,516, filed Dec. 9, 2019, which is also a continuation-in-part ofSer. No. 16/123,735, filed Sep. 6, 2018, now U.S. Pat. No. 10,765,117,which is a continuation of Ser. No. 15/445,247, filed Feb. 28, 2017, nowU.S. Pat. No. 10,070,649, which is a continuation-in-part of U.S. patentapplication Ser. No. 14/690,929, filed on Apr. 20, 2015, now U.S. Pat.No. 9,585,396, which claims the benefit of priority under 35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 61/991,821, filed May12, 2014, the contents of all of which are incorporated by reference intheir entireties.

U.S. patent application Ser. No. 14/690,929, now U.S. Pat. No.9,585,396, filed on Apr. 20, 2015, is also a continuation-in-part ofU.S. patent application Ser. No. 14/294,057, now U.S. Pat. No.9,426,996, filed on Jun. 2, 2014, which is a continuation of U.S. patentapplication Ser. No. 14/167,093, now U.S. Pat. No. 9,138,001, filed onJan. 29, 2014, which claims the benefit of priority under 35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 61/758,313, filed Jan.30, 2013, the contents of all of which are hereby incorporated byreference in their entireties.

U.S. patent application Ser. No. 14/690,929, now U.S. Pat. No.9,585,396, filed on Apr. 20, 2015, is also a continuation-in-part ofU.S. patent application Ser. No. 14/182,793, now U.S. Pat. No.9,138,002, filed on Feb. 18, 2014, which is a divisional of U.S. patentapplication Ser. No. 13/945,577, now U.S. Pat. No. 8,669,207, filed onJul. 18, 2013, which claims the benefit of priority under 35 U.S.C. §119(e) of 61/831,187, filed Jun. 5, 2013, and U.S. Provisional PatentApplication No. 61/758,313, filed Jan. 30, 2013, the contents of all ofwhich are hereby incorporated by reference in their entireties.

FIELD OF THE PRESENT APPLICATION

The present application relates to large-scale methods of uniformlycoating packaging surfaces with a benzoxaborole compound.

BACKGROUND

Benzoxaborole is a drug known to be effective in treating eukaryoticfungal and parasitic infections. For example, benzoxaborole is used totreat fungal conditions affecting the toenails and fingernails ofhumans, such as Onychomycosis. Benzoxaborole is also known to be aneffective treatment of Human African Trypanosomiasis, commonly calledAfrican Sleeping Sickness, which is caused by T. brucei parasites thatinfect thousands of people annually in sub-Saharan Africa.

Benzoxaborole has also been shown to have antimicrobial effects inplants. For example, benzoxaborole compounds have been proven to beeffective as a volatile plant fungicide. However, coating packagingsurfaces with a volatile biological control agent, such as abenzoxaborole compound, to provide antimicrobial protection to food,such as crops and meats, and to preserve food freshness has not beenreported.

The present disclosure describes large-scale methods of applyingvolatile antimicrobial compounds on food containers and packagingmaterials to inhibit microorganisms responsible for decay. Morespecifically, the present disclosure provides methods of coatingbenzoxaboroles on the surface of food containers and packaging materialsin mass in order to provide antimicrobial protection to food. Inaddition, the present disclosure describes methods to treat foodproducts against microorganisms that are detrimental to the preservationof the freshness of food, such as plant, crops, or meats.

SUMMARY OF THE INVENTION

The present disclosure provides a method of treating food products withan antimicrobial agent. The method comprises administering anantimicrobial agent, sometimes called a benzoxaborole compound, to oneor more surfaces of a chamber. The method also comprises drying the oneor more surfaces of the chamber. The method further comprises placing afood product inside of the chamber. Finally, the method comprisesvaporizing the benzoxaborole compound from the one or more surfaces ofthe chamber to treat the food product.

In the method described herein, the food product may be a plant crop ora meat, wherein the crop may be a fruit selected from the groupconsisting of a strawberry, a raspberry, a blackberry, or a blueberry.In addition, the benzoxaborole compound of the present method may beselected from the group consisting of Compound A, Compound B, and/orCompound C. The chamber of the method may be a clamshell, and theclamshell may comprise polyethylene terephthalate. Finally, thebenzoxaborole compound of the present method may be administered to thechamber by drenching, spraying, painting, vaporizing, and/orsublimation.

The present disclosure is also directed to a large-scale method oftreating a plurality of chambers with an antimicrobial agent. The methodcomprises placing a plurality of chambers in a position to be treated,wherein each of the plurality of chambers comprises one or moresurfaces. The method further comprises administering a benzoxaborolecompound to the one or more surfaces of the plurality of chambers. Inaddition, the method comprises drying the one or more surfaces of theplurality of chambers, wherein the drying may be instantaneous. Further,the large-scale method comprises affixing the benzoxaborole compound tothe one or more surfaces of the plurality of chambers.

The benzoxaborole compound of the present large-scale method may beselected from the group consisting of Compound A, Compound B, and/orCompound C, and may be administered to the plurality of chambers bydrenching, spraying, painting, vaporizing, or sublimation.

The benzoxaborole compound may also be administered to the plurality ofchambers during preformation, formation, or postformation of theplurality of chambers. The surfaces of the plurality of chambers of thelarge-scale method may further comprise a liquid-absorbing material. Theplurality of chambers of the large-scale method may comprise a pluralityof clamshells, and the plurality of clamshells may comprise polyethyleneterephthalate.

DETAILED DESCRIPTION

The following numbered embodiments are contemplated and arenon-limiting:

1. A method of treating a food product with an antimicrobial agent, themethod comprising:

administering a benzoxaborole treatment to one or more surfaces of afood packaging material, wherein the benzoxaborole treatment comprisesone or more benzoxaborole compounds,

drying the one or more surfaces of the food packaging material,

placing a food product inside of the food packaging material, and

vaporizing the benzoxaborole compound from the one or more surfaces ofthe food packaging material to treat the food product located therein.

2. The method of clause 1, wherein the food product is a plant, a crop,or a meat.

3. The method of clause 1 or clause 2, wherein the food product is afruit.

4. The method of any of clauses 1 to 3, wherein the food product isselected from the group consisting of a strawberry, a raspberry, ablackberry, and a blueberry.

5. The method of any one of clauses 1 to 4, wherein the benzoxaborolecompound is selected from the group consisting of Compound A, CompoundB, Compound C, and combinations thereof.

6. The method of any one of clauses 1 to 5, wherein Compound A has thestructure:

or an analog or a derivative thereof.

7. The method of any one of clauses 1 to 5, wherein Compound B has thestructure:

or an analog or a derivative thereof.

8. The method of any one of clauses 1 to 5, wherein Compound C has thestructure:

or an analog or a derivative thereof.

9. The method of any one of clauses 1 to 8, wherein the benzoxaborolecompound is in the form of a liquid, a gas, or a solid.

10. The method of any one of clauses 1 to 9, wherein the food packagingmaterial is comprised by a chamber.

11. The method of clause 10, wherein the chamber is selected from thegroup consisting of a container, a liner material, one chamber, and aplurality of chambers.

12. The method of clause 10 or clause 11, wherein the chamber is aclamshell.

13. The method of clause 12, wherein the clamshell comprisespolyethylene terephthalate (PET).

14. The method of any one of clauses 1 to 13, wherein the benzoxaboroletreatment is embedded into, impregnated within, or coated onto the foodpackaging material.

15. The method of any one of clauses 1 to 14, wherein the benzoxaboroletreatment is embedded into the food packaging material.

16. The method of any one of clauses 1 to 14, wherein the benzoxaboroletreatment is coated onto the food packaging material.

17. The method of any one of clauses 1 to 14, wherein the benzoxaboroletreatment is impregnated into the food packaging material.

18. The method of any one of clauses 1 to 17, wherein the benzoxaboroletreatment is in the form of a spray, a liquid, a mist, a gel, a thermalfog, a non-thermal fog, a dip, a drench, a vapor, a gas, or sublimation.

19. The method of any one of clauses 1 to 18, wherein the benzoxaboroletreatment further comprises a treatment carrier.

20. The method of clause 19, wherein the treatment carrier comprises aliquid, a gas, a solution, a solvent, and a chemical.

21. The method of clause 19 or clause 20, wherein the treatment carrieris a liquid.

22. The method of any one of clauses 19 to 21, wherein the treatmentcarrier is selected from the group consisting of water, saline, abuffer, a solution, a solvent, a solvent-based solution, and acombination thereof.

23. The method of clause any one of clauses 19 to 22, wherein thetreatment carrier is supercritical CO₂.

24. The method of clause 19 or clause 20, wherein the treatment carrieris a gas.

25. The method of any one of clauses 19, 20, or 24, wherein thetreatment carrier is selected from the group consisting of nitrogen(N₂), carbon dioxide (CO₂), and sulfur dioxide (SO₂).

26. The method of any one of clauses 19, 20, 24, or 25, wherein thetreatment carrier is nitrogen (N₂).

27. The method of any one of clauses 19, 20, 24, or 25, wherein thetreatment carrier is carbon dioxide (CO₂).

28. The method of any one of clauses 19, 20, 24, or 25, wherein thetreatment carrier is sulfur dioxide (SO₂).

29. The method of any one of clauses 1 to 28, wherein the benzoxaboroletreatment is effective against plant pathogens.

30. The method of clause 29, wherein the plant pathogens are fungalpathogens.

31. The method of clause 29 or clause 30, wherein the plant pathogensare selected from the group consisting of Acremonium spp., Albugo spp.,Alternaria spp., Ascochyta spp., Aspergillus spp., Botryodiplodia spp.,Botryospheria spp., Botrytis spp., Byssochlamys spp., Candida spp.,Cephalosporium spp., Ceratocystis spp., Cercospora spp., Chalara spp.,Cladosporium spp., Colletotrichum spp., Cryptosporiopsis spp.,Cylindrocarpon spp., Debaryomyces spp., Diaporthe spp., Didymella spp.,Diplodia spp., Dothiorella spp., Elsinoe spp., Fusarium spp., Geotrichumspp., Gloeosporium spp., Glomerella spp., Helminthosporium spp., Khuskiaspp., Lasiodiplodia spp., Macrophoma spp., Macrophomina spp.,Microdochium spp., Monilinia spp., Monilochaethes spp., Mucor spp.,Mycocentrospora spp., Mycosphaerella spp., Nectria spp., Neofabraeaspp., Nigrospora spp., Penicillium spp., Peronophythora spp.,Peronospora spp., Pestalotiopsis spp., Pezicula spp., Phacidiopycnisspp., Phoma spp., Phomopsis spp., Phyllosticta spp., Phytophthora spp.,Polyscytalum spp., Pseudocercospora spp., Pyricularia spp., Pythiumspp., Rhizoctonia spp., Rhizopus spp., Sclerotium spp., Sclerotiniaspp., Septoria spp., Sphaceloma spp., Sphaeropsis spp., Stemphylliumspp., Stilbella spp., Thielaviopsis spp., Thyronectria spp.,Trachysphaera spp., Uromyces spp., Ustilago spp., Venturia spp., andVerticillium spp., and bacterial pathogens, such as Bacillus spp.,Campylobacter spp., Clavibacter spp., Clostridium spp., Erwinia spp.,Escherichia spp., Lactobacillus spp., Leuconostoc spp., Listeria spp.,Pantoea spp., Pectobacterium spp., Pseudomonas spp., Ralstonia spp.,Salmonella spp., Shigella spp., Staphylococcus spp., Vibrio spp.,Xanthomonas spp., and Yersinia spp.

32. The method of any one of clauses 29 to 31, wherein the plantpathogens are selected from the group consisting of Botrytis cinerea,Mucor piriformis, Fusarium sambucinum, Aspergillus brasiliensis, andPeniciliium expansum.

33. The method of any one of clauses 10 to 32, wherein the chamber isopen, closed, or sealed.

34. The method of any one of clauses 10 to 33, wherein the chamber issealed.

35. The method of any one of clauses 10 to 34, wherein the chamber isair-tight.

36. The method of any one of clauses 10 to 35, wherein the chamber issemipermeable or impermeable.

37. The method of any one of clauses 10 to 36, wherein the chamber ismade of a material selected from the group consisting of cardboard,paper, paperboard, corrugated paper, plastic, glass, polyester,polystyrene, cellulosic material, metal, and cement.

38. The method of clause 37, wherein the metal is selected from thegroup consisting of aluminum, foils, laminates, tinplate, and steel.

39. The method of clause 38, wherein the steel is tin-free steel.

40. The method of clause 37, wherein the plastic is selected from thegroup consisting of thermosets and thermoplastics.

41. The method of clause 37, wherein the polyester is selected from thegroup consisting of polycarbonate, polyethylene naphthalate, andpolyethylene terephthalate (PET).

42. The method of any one of clauses 10 to 41, wherein the chambercomprises a port, an outlet, or both.

43. The method of any one of clauses 10 to 42, wherein the chamber has avolume from between about 0.1 L to about 50 L.

44. The method of any one of clauses 10 to 43, wherein the chambercomprises a plurality of individual clamshells.

45. The method of any one of clauses 10 to 44, wherein the chambercomprises between about 2 clamshells to about 384,000,000 clamshells.

46. The method of any one of clauses 10 to 45, wherein the chamberfurther comprises a liquid-absorbing material.

47. The method of clause 46, wherein the liquid-absorbing material iscomprised on the interior of the chamber or on the exterior of thechamber.

48. The method of clause 46 or clause 47, wherein the liquid-absorbingmaterial is comprised on the interior of the chamber.

49. The method of clause 47 or clause 48, wherein the liquid-absorbingmaterial is comprised on the internal top, the internal bottom, or theinternal side panels of the chamber.

50. The method of clause 46 or clause 47, wherein the liquid-absorbingmaterial is comprised on the exterior of the chamber.

51. The method of clause 47 or clause 50, wherein the liquid-absorbingmaterial is comprised on the external top, the external bottom, or theexternal side panels of the chamber.

52. The method of any one of clauses 46 to 51, wherein theliquid-absorbing material is attached or affixed to the chamber.

53. The method of any one of clauses 46 to 52, wherein theliquid-absorbing material is comprised in a chamber component selectedfrom the group consisting of a liner, a wrapping, a label, a tag, asticker, and a pad.

54. The method of any one of clauses 45 to 53, wherein theliquid-absorbing material is selected from the group consisting ofcotton, paper, and foam.

55. The method of any one of clauses 46 to 54, wherein theliquid-absorbing material is a reservoir capable of releasing thebenzoxaborole treatment to the food product comprised in the chamber.

56. The method of any one of clauses 46 to 55, wherein theliquid-absorbing material provides for quick-release or slow-release ofthe benzoxaborole treatment to the food product comprised in the chamberover a time period.

57. The method of any one of clauses 46 to 56, wherein theliquid-absorbing material provides for quick-release of thebenzoxaborole treatment to the food product comprised in the chamberover a time period.

58. The method of any one of clauses 46 to 56, wherein theliquid-absorbing material provides for slow-release of the benzoxaboroletreatment to the food product comprised in the chamber over a timeperiod.

59. The method of clause 56 or clause 57, wherein the time period forquick-release of the benzoxaborole treatment by the liquid-absorbingmaterial is about 12 hours or less.

60. The method of any one of clauses 56, 57, or 59, wherein the timeperiod for quick-release of the benzoxaborole treatment by theliquid-absorbing material is between about 5 seconds to about 12 hoursor less.

61. The method of clause 56 or clause 58, wherein the time period forslow-release of the benzoxaborole treatment by the liquid-absorbingmaterial is over 12 hours.

62. The method of any one of clauses 56, 58, or 61, wherein the timeperiod for slow-release of the benzoxaborole treatment by theliquid-absorbing material is between over 12 hours to about 31 days.

63. The method of any one of clauses 10 to 62, wherein the chamberfurther comprises one or more apertures.

64. The method of clause 63, wherein the one or more apertures has asize between about 2 mm to about 2 cm.

65. The method of clause 63 or clause 64, wherein the one or moreapertures has a location on the chamber selected from the groupconsisting of the base, the lid, the sides, or a combination thereof.

66. The method of any one of clauses 63 to 65, wherein the one or moreapertures enables introduction of the benzoxaborole treatment, thetreatment carrier, or a combination thereof to the chamber.

67. The method of any one of clauses 63 to 65, wherein the one or moreapertures enables release of the benzoxaborole treatment, the treatmentcarrier, or a combination thereof from the chamber.

68. The method of any one of clauses 1 to 67, wherein the food productsare manually or robotically placed in the chamber.

69. The method of any one of clauses 1 to 68, wherein the food productsare treated post-harvest.

70. The method of any one of clauses 1 to 69, wherein the distancebetween the food products and the food packaging material comprising thebenzoxaborole treatment is no greater than 6 feet.

71. The method of any one of clauses 1 to 70, wherein the distancebetween the food products and the food packaging material comprising thebenzoxaborole treatment is between about 0.1 inches and about 6 feet.

72. The method any one of clauses 1 to 71, wherein the one or more foodproducts are treated for an initial time period ranging from about 12hours to about 5 days.

73. The method of any one of clauses 1 to 72, wherein the benzoxaboroletreatment concentration ranges from about 0.1 mg/chamber to about 10mg/chamber.

74. The method of any one of clauses 1 to 73, wherein drying the one ormore surfaces of the food packaging material occurs at room temperature,wherein room temperature is between about 21° C. and about 23° C.

75. The method of any one of clauses 1 to 74, wherein drying the one ormore surfaces of the food packaging material occurs instantaneously orwithin seconds.

76. The method of any one of clauses 1 to 75, wherein drying the one ormore surfaces of the food packaging material occurs instantaneously.

77. The method of any one of clauses 1 to 75, wherein drying the one ormore surfaces of the food packaging material occurs within seconds

78. The method of any one of clauses 1 to 75 or 77, wherein drying theone or more surfaces of the food packaging material occurs between about0.1 seconds and about 60 seconds.

79. The method of any one of clauses 1 to 78, wherein the method resultsin greater uniformity and consistency in application of benzoxaboroletreatment to the food packaging material.

80. The method of any one of clauses 1 to 79, wherein the methodprovides one month of extended antimicrobial protection to the treatedfood products.

A large-scale method of treating a plurality of chambers with an

antimicrobial agent, the method comprising:

placing a plurality of chambers in a position to be treated wherein eachof the chambers comprise one or more surfaces,

administering the benzoxaborole treatment to one or more surfaces of theplurality of chambers during preformation, formation, or postformationof the plurality of chambers, wherein the benzoxaborole treatmentcomprises one or more benzoxaborole compounds,

drying the one or more surfaces of the plurality of chambers, and

affixing the benzoxaborole compound to the one or more surfaces of the

plurality of chambers.

The method of clause 81, wherein the plurality of chambers further

comprise a food product that is a plant, a crop, or a meat.

The method of clause 82, wherein the food product is a fruit.

The method of any of clauses 82 or clause 83, wherein the food product

is selected from the group consisting of a strawberry, a raspberry, ablackberry, and a blueberry.

The method of any one of clauses 81 to 84, wherein the benzoxaborole

compound is selected from the group consisting of Compound A, CompoundB, Compound C, and combinations thereof.

86. The method of any one of clauses 81 to 85, wherein Compound A hasthe structure:

or an analog or a derivative thereof.

87. The method of any one of clauses 81 to 85, wherein Compound B hasthe structure:

or an analog or a derivative thereof.

88. The method of any one of clauses 81 to 85, wherein Compound C hasthe structure:

or an analog or a derivative thereof.

89. The method of any one of clauses 81 to 88, wherein the benzoxaborolecompound is in the form of a liquid, a gas, or a solid.

90. The method of any one of clauses 81 to 89, wherein the plurality ofchambers comprise one or more containers.

91. The method of any one of clauses 81 to 90, wherein the plurality ofchambers comprise one or more liner materials.

92. The method of any one of clauses 81 to 91, wherein the plurality ofchambers comprise one or more clamshells.

93. The method of any one of clauses 81 to 92, wherein the clamshellscomprise polyethylene terephthalate (PET).

94. The method of any one of clauses 81 to 93, wherein the benzoxaboroletreatment is embedded into, impregnated within, or coated onto theplurality of chambers.

95. The method of any one of clauses 81 to 94, wherein the benzoxaboroletreatment is embedded into the plurality of chambers.

96. The method of any one of clauses 81 to 94, wherein the benzoxaboroletreatment is coated onto the plurality of chambers.

97. The method of any one of clauses 81 to 94, wherein the benzoxaboroletreatment is impregnated into the plurality of chambers.

98. The method of any one of clauses 81 to 97, wherein the benzoxaboroletreatment is in the form of a spray, a liquid, a mist, a gel, a thermalfog, a non-thermal fog, a dip, a drench, a vapor, a gas, or sublimation.

99. The method of any one of clauses 81 to 98, wherein the benzoxaboroletreatment further comprises a treatment carrier.

100. The method of clause 99, wherein the treatment carrier comprises aliquid, a gas, a solution, a solvent, and a chemical.

101. The method of clause 99 or clause 100, wherein the treatmentcarrier is a liquid.

102. The method of any one of clauses 99 to 101, wherein the treatmentcarrier is selected from the group consisting of water, saline, abuffer, a solution, a solvent, a solvent-based solution, and acombination thereof.

103. The method of clause any one of clauses 99 to 102, wherein thetreatment carrier is supercritical CO₂.

104. The method of clause 99 or clause 100, wherein the treatmentcarrier is a gas.

105. The method of any one of clauses 99, 100, or 104, wherein thetreatment carrier is selected from the group consisting of nitrogen(N₂), carbon dioxide (CO₂), and sulfur dioxide (SO₂).

106. The method of any one of clauses 99, 100, 104, or 105, wherein thetreatment carrier is nitrogen (N₂).

107. The method of any one of clauses 99, 100, 104, or 105, wherein thetreatment carrier is carbon dioxide (CO₂).

108. The method of any one of clauses 99, 100, 104, or 105, wherein thetreatment carrier is sulfur dioxide (SO₂).

109. The method of any one of clauses 81 to 108, wherein thebenzoxaborole treatment is effective against plant pathogens.

110. The method of clause 109, wherein the plant pathogens are fungalpathogens.

111. The method of clause 109 or clause 110, wherein the plant pathogensare selected from the group consisting of Acremonium spp., Albugo spp.,Alternaria spp., Ascochyta spp., Aspergillus spp., Botryodiplodia spp.,Botryospheria spp., Botrytis spp., Byssochlamys spp., Candida spp.,Cephalosporium spp., Ceratocystis spp., Cercospora spp., Chalara spp.,Cladosporium spp., Colletotrichum spp., Cryptosporiopsis spp.,Cylindrocarpon spp., Debaryomyces spp., Diaporthe spp., Didymella spp.,Diplodia spp., Dothiorella spp., Elsinoe spp., Fusarium spp., Geotrichumspp., Gloeosporium spp., Glomerella spp., Helminthosporium spp., Khuskiaspp., Lasiodiplodia spp., Macrophoma spp., Macrophomina spp.,Microdochium spp., Monilinia spp., Monilochaethes spp., Mucor spp.,Mycocentrospora spp., Mycosphaerella spp., Nectria spp., Neofabraeaspp., Nigrospora spp., Penicillium spp., Peronophythora spp.,Peronospora spp., Pestalotiopsis spp., Pezicula spp., Phacidiopycnisspp., Phoma spp., Phomopsis spp., Phyllosticta spp., Phytophthora spp.,Polyscytalum spp., Pseudocercospora spp., Pyricularia spp., Pythiumspp., Rhizoctonia spp., Rhizopus spp., Sclerotium spp., Sclerotiniaspp., Septoria spp., Sphaceloma spp., Sphaeropsis spp., Stemphylliumspp., Stilbella spp., Thielaviopsis spp., Thyronectria spp.,Trachysphaera spp., Uromyces spp., Ustilago spp., Venturia spp., andVerticillium spp., and bacterial pathogens, such as Bacillus spp.,Campylobacter spp., Clavibacter spp., Clostridium spp., Erwinia spp.,Escherichia spp., Lactobacillus spp., Leuconostoc spp., Listeria spp.,Pantoea spp., Pectobacterium spp., Pseudomonas spp., Ralstonia spp.,Salmonella spp., Shigella spp., Staphylococcus spp., Vibrio spp.,Xanthomonas spp., and Yersinia spp.

112. The method of any one of clauses 109 to 111, wherein the plantpathogens are selected from the group consisting of Botrytis cinerea,Mucor piriformis, Fusarium sambucinum, Aspergillus brasiliensis, andPeniciliium expansum.

113. The method of any one of clauses 90 to 112, wherein the chamber isopen, closed, or sealed.

114. The method of clause 113, wherein the chamber is sealed.

115. The method of clause 113 or clause 114, wherein the chamber issealed air-tight.

116. The method of any one of clauses 90 to 115, wherein the chamber issemipermeable or impermeable.

117. The method of any one of clauses 90 to 116, wherein the chamber ismade of a material selected from the group consisting of cardboard,paper, paperboard, corrugated paper, plastic, glass, polyester,polystyrene, cellulosic material, metal, and cement.

118. The method of clause 117, wherein the metal is selected from thegroup consisting of aluminum, foils, laminates, tinplate, and steel.

119. The method of clause 118, wherein the steel is tin-free steel.

120. The method of clause 117, wherein the plastic is selected from thegroup consisting of thermosets and thermoplastics.

121. The method of clause 117, wherein the polyester is selected fromthe group consisting of polycarbonate, polyethylene naphthalate, andpolyethylene terephthalate (PET).

122. The method of any one of clauses 90 to 121, wherein the chambercomprises a port, an outlet, or both.

123. The method of any one of clauses 81 to 122, wherein the chamber hasa volume from between about 0.1 L to about 50 L.

124. The method of any one of clauses 81 to 123, wherein the chambercomprises a plurality of individual clamshells.

125. The method of any one of clauses 81 to 124, wherein the chambercomprises between about 2 clamshells to about 384,000,000 clamshells.

126. The method of any one of clauses 81 to 125, wherein the chamberfurther comprises a liquid-absorbing material.

127. The method of clause 126, wherein the liquid-absorbing material iscomprised on the interior of the chamber or on the exterior of thechamber.

128. The method of clause 126 or clause 127, wherein theliquid-absorbing material is comprised on the interior of the chamber.

129. The method of clause 127 or clause 128, wherein theliquid-absorbing material is comprised on the internal top, the internalbottom, or the internal side panels of the chamber, or combinationsthereof.

130. The method of clause 126 or clause 127, wherein theliquid-absorbing material is comprised on the exterior of the chamber.

131. The method of clause 127 or clause 130, wherein theliquid-absorbing material is comprised on the external top, the externalbottom, or the external side panels of the chamber, or combinationsthereof.

132. The method of any one of clauses 126 to 131, wherein theliquid-absorbing material is attached or affixed to the chamber.

133. The method of any one of clauses 126 to 132, wherein theliquid-absorbing material is comprised in a chamber component selectedfrom the group consisting of a liner, a wrapping, a label, a tag, asticker, and a pad.

134. The method of any one of clauses 126 to 133, wherein theliquid-absorbing material is selected from the group consisting ofcotton, paper, and foam.

135. The method of any one of clauses 126 to 134, wherein theliquid-absorbing material is a reservoir capable of releasing thebenzoxaborole treatment to the food product comprised in the chamber.

136. The method of any one of clauses 126 to 135, wherein theliquid-absorbing material provides for quick-release or slow-release ofthe benzoxaborole treatment to the food product comprised in the chamberover a time period.

137. The method of any one of clauses 126 to 136, wherein theliquid-absorbing material provides for quick-release of thebenzoxaborole treatment to the food product comprised in the chamberover a time period.

138. The method of any one of clauses 126 to 136, wherein theliquid-absorbing material provides for slow-release of the benzoxaboroletreatment to the food product comprised in the chamber over a timeperiod.

139. The method of clause 136 or clause 137, wherein the time period forquick-release of the benzoxaborole treatment by the liquid-absorbingmaterial is about 12 hours or less.

140. The method of any one of clauses 136, 137, or 139, wherein the timeperiod for quick-release of the benzoxaborole treatment by theliquid-absorbing material is between about 5 seconds to about 12 hoursor less.

141. The method of clause 136 or clause 138, wherein the time period forslow-release of the benzoxaborole treatment by the liquid-absorbingmaterial is over 12 hours.

142. The method of any one of clauses 136, 138, or 141, wherein the timeperiod for slow-release of the benzoxaborole treatment by theliquid-absorbing material is between over 12 hours to about 31 days.

143. The method of any one of clauses 81 to 142, wherein the pluralityof chambers further comprises one or more apertures.

144. The method of clause 143, wherein the one or more apertures has asize between about 2 mm to about 2 cm.

145. The method of clause 143 or clause 144, wherein the one or moreapertures is in a location on the chamber selected from the groupconsisting of the base, the lid, the sides, or a combination thereof.

146. The method of any one of clauses 143 to 145, wherein the one ormore apertures enables introduction of the benzoxaborole treatment, thetreatment carrier, or a combination thereof to the chamber.

147. The method of any one of clauses 143 to 145, wherein the one ormore apertures enables release of the benzoxaborole treatment, thetreatment carrier, or a combination thereof from the chamber.

148. The method of any one of clauses 81 to 147, wherein the foodproducts are manually or robotically placed in the chamber.

149. The method of any one of clauses 81 to 148, wherein the foodproducts are treated post-harvest.

150. The method of any one of clauses 81 to 149, wherein the distancebetween the food products and the plurality of chambers comprising thebenzoxaborole treatment is no greater than 6 feet.

151. The method of any one of clauses 81 to 150, wherein the distancebetween the food products and the plurality of chambers comprising thebenzoxaborole treatment is between about 0.1 inches and about 6 feet.

152. The method any one of clauses 81 to 151, wherein the one or morefood products are treated for an initial time period ranging from about12 hours to about 5 days.

153. The method of any one of clauses 81 to 152, wherein thebenzoxaborole treatment concentration ranges from about 0.1 mg/chamberto about 10 mg/chamber.

154. The method of any one of clauses 81 to 153, wherein drying the oneor more surfaces of the plurality of chambers occurs at roomtemperature, wherein room temperature is between about 21° C. and about23° C.

155. The method of any one of clauses 81 to 154, wherein drying the oneor more surfaces of the plurality of chambers occurs instantaneously orwithin seconds.

156. The method of any one of clauses 81 to 155, wherein drying the oneor more surfaces of the plurality of chambers occurs instantaneously.

157. The method of any one of clauses 81 to 155, wherein drying the oneor more surfaces of the plurality of chambers occurs within seconds.

158. The method of any one of clauses 81 to 155 or 157, wherein dryingthe one or more surfaces of the plurality of chambers occurs betweenabout 0.1 seconds and about 60 seconds.

159. The method of any one of clauses 81 to 158, wherein the methodresults in greater uniformity and consistency in application ofbenzoxaborole treatment to the plurality of chambers.

160. The method of any one of clauses 81 to 159, wherein the methodprovides one month of extended antimicrobial protection to the treatedfood products.

The terms “chamber,” “container,” “material,” or the phrase “packagingmaterial” are interchangeable and refer to any material that is used tobox, wrap, store, or package a food or food product, such as a plant,crop, or meat. A plurality of chambers comprises from about 1000 to tensof thousands to one or more millions of chambers.

The term “carrier” refers to a material, composition, or control, suchas a liquid or solid filler, diluent, excipient, solvent, gas, orencapsulating material, involved in carrying or transporting an activeingredient, compound, analog, or derivative from one location to anotherlocation. A carrier must be acceptable in the sense of being compatiblewith the other ingredients of the formulation and not injurious to food,such as plant, crops, or meat products.

Exemplary embodiments of the compounds of the present disclosure aredescribed herein. In some embodiments, the antimicrobial compound,sometimes called a benzoxaborole, comprises Compounds A, B, and/or C,which may encompass diastereomers and enantiomers of the illustrativecompounds. Enantiomers are defined as one of a pair of molecularentities which are mirror images of each other and non-superimposable.Diastereomers or diastereoisomers are defined as stereoisomers otherthan enantiomers. Diastereomers or diastereoisomers are stereoisomersnot related as mirror images. Diastereoisomers are characterized bydifferences in physical properties.

Unless otherwise stated, the following terms used in this application,including the specification and claims, have the definitions givenbelow. It must be noted that, as used in the specification and theappended claims, the singular forms “a,” “an” and “the” include pluralreferents unless the context clearly dictates otherwise. Definition ofstandard chemistry terms may be found in reference works, includingCarey and Sundberg, Advanced Organic Chemistry 4^(th) Ed., Vols. A(2000) and B (2001), Plenum Press, New York, N.Y.

As used herein, the phrase “moiety” refers to a specific segment orfunctional group of a molecule. Chemical moieties are often recognizedchemical entities embedded in or appended to a molecule.

As used herein, the phrases “heteroatom” and “hetero-” refer to atomsother than carbon (C) and hydrogen (H). Examples of heteroatoms includeoxygen (O), nitrogen (N) sulfur (S), silicon (Si), germanium (Ge),aluminum (Al) and boron (B).

As used herein, the phrases “halo” and “halogen” are interchangeable andrefer to fluoro (—F), chloro (—Cl), bromo (—Br), and iodo (—I).

As used herein, the phrase “alkyl” refers to an unsubstituted orsubstituted, hydrocarbon group and can include straight, branched,cyclic, saturated and/or unsaturated features. Although the alkyl moietymay be an “unsaturated alkyl” moiety, which means that it contains atleast one alkene or alkyne moiety, typically, the alkyl moiety is a“saturated alkyl” group, which means that it does not contain any alkeneor alkyne moieties. Likewise, although the alkyl moiety may be cyclic,the alkyl moiety typically is acyclic group. Thus, in some embodiments,“alkyl” refers to an optionally substituted straight-chain, oroptionally substituted branched-chain saturated hydrocarbon monoradicalhaving from about one to about thirty carbon atoms in some embodiments,from about one to about fifteen carbon atoms in some embodiments, andfrom about one to about six carbon atoms in further embodiments.Examples of saturated alkyl radicals include, but are not limited to,methyl, ethyl, n-propyl, isopropyl, 2-methyl-1-propyl,2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl,2,2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl,4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl,4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl,2-ethyl-1-butyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl,isopentyl, neopentyl, and n-hexyl, and longer alkyl groups, such asheptyl, and octyl. It should be noted that whenever it appears herein, anumerical range such as “1 to 6” refers to each integer in the givenrange; e.g., “1 to 6 carbon atoms” or “C₁₋₆” or “C₁-C₆” means that thealkyl group may consist of 1 carbon atom, 2 carbon atoms, 3 carbonatoms, 4 carbon atoms, 5 carbon atoms, and/or 6 carbon atoms, althoughthe present definition also covers the occurrence of the term “alkyl”where no numerical range is designated.

As used herein, the phrase “substituted alkyl” refers to an alkyl group,as defined herein, in which one or more (up to about five, preferably upto about three) hydrogen atoms is replaced by a substituentindependently selected from the substituent group defined herein.

As used herein, the phrases “substituents” and “substituted” refer togroups which may be used to replace another group on a molecule. Suchgroups are known to those of skill in the chemical arts and may include,without limitation, one or more of the following independently selectedgroups, or designated subsets thereof: halogen, —CN, —OH, —NO₂, —N₃, ═O,═S, ═NH, —SO₂, —NH₂, —COOH, nitroalkyl, amino, including mono- anddi-substituted amino groups, cyanato, isocyanato, thiocyanato,isothiocyanato, guanidinyl, O-carbamyl, N-carbamyl, thiocarbamyl, uryl,isouryl, thiouryl, isothiouryl, mercapto, sulfanyl, sulfinyl, sulfonyl,sulfonamidyl, phosphonyl, phosphatidyl, phosphoramidyl, dialkylamino,diarylamino, diarylalkylamino; and the protected compounds thereof. Theprotecting groups that may form the protected compounds of the abovesubstituents are known to those of skill in the art and may be found inreferences such as Greene and Wuts, Protective Groups in OrganicSynthesis, 3rd ed.; John Wiley & Sons, New York, N.Y. (1999) andKocienski, Protective Groups; Thieme Verlag, New York, N.Y. (1994) whichare incorporated herein by reference in their entirety.

As used herein, the phrase “alkoxy” refers to the group —O-alkyl, wherealkyl is as defined herein. In one embodiment, alkoxy groups include,e.g., methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy,sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like. Thealkoxy can be unsubstituted or substituted.

As used herein, the phrases “cyclic” and “membered ring” refer to anycyclic structure, including alicyclic, heterocyclic, aromatic,heteroaromatic and polycyclic fused or non-fused ring systems asdescribed herein. The term “membered” is meant to denote the number ofskeletal atoms that constitute the ring. Thus, for example, pyridine,pyran, and pyrimidine are six-membered rings and pyrrole,tetrahydrofuran, and thiophene are five-membered rings.

As used herein, the phrase “aromatic” refers to a cyclic or polycyclicmoiety having a conjugated unsaturated (4n+2)π electron system (where nis a positive integer), sometimes referred to as a delocalized πelectron system.

As used herein, the phrase “aryl” refers to an optionally substituted,aromatic, cyclic, hydrocarbon monoradical of from six to about twentyring atoms, preferably from six to about ten carbon atoms and includesfused (or condensed) and non-fused aromatic rings. A fused aromatic ringradical contains from two to four fused rings where the ring ofattachment is an aromatic ring, and the other individual rings withinthe fused ring may be cycloalkyl, cycloalkenyl, cycloalkynyl,heterocycloalkyl, heterocycloalkenyl, heterocycloalkynyl, aromatic,heteroaromatic or any combination thereof. A non-limiting example of asingle ring aryl group includes phenyl; a fused ring aryl group includesnaphthyl, anthryl, azulenyl; and a non-fused bi-aryl group includesbiphenyl.

As used herein, the phrase “substituted aryl” refers to an aryl group,as defined herein, in which one or more (up to about five, preferably upto about three) hydrogen atoms is replaced by a substituentindependently selected from the group defined herein, (except asotherwise constrained by the definition for the aryl substituent).

As used herein, the phrase “heteroaryl” refers to an optionallysubstituted, aromatic, cyclic monoradical containing from about five toabout twenty skeletal ring atoms, preferably from five to about ten ringatoms and includes fused (or condensed) and non-fused aromatic rings,and which have one or more (one to ten, preferably about one to aboutfour) ring atoms selected from an atom other than carbon (i.e., aheteroatom) such as, for example, oxygen, nitrogen, sulfur, selenium,phosphorus or combinations thereof. The term heteroaryl includesoptionally substituted fused and non-fused heteroaryl radicals having atleast one heteroatom. A fused heteroaryl radical may contain from two tofour fused rings where the ring of attachment is a heteroaromatic ringand the other individual rings within the fused ring system may bealicyclic, heterocyclic, aromatic, heteroaromatic or any combinationthereof. The term heteroaryl also includes fused and non-fusedheteroaryls having from five to about twelve skeletal ring atoms, aswell as those having from five to about ten skeletal ring atoms.Examples of heteroaryl groups include, but are not limited to,acridinyl, benzo[1,3]dioxole, benzimidazolyl, benzindazolyl,benzoisooxazolyl, benzokisazolyl, benzofuranyl, benzofurazanyl,benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzo[b]thienyl,benzothiophenyl, benzothiopyranyl, benzotriazolyl, benzoxazolyl,carbazolyl, carbolinyl, chromenyl, cinnolinyl, furanyl, furazanyl,furopyridinyl, furyl, imidazolyl, indazolyl, indolyl, indolidinyl,indolizinyl, isobenzofuranyl, isoindolyl, isoxazolyl, isoquinolinyl,isothiazolyl, naphthylidinyl, naphthyridinyl, oxadiazolyl, oxazolyl,phenoxazinyl, phenothiazinyl, phenazinyl, phenoxathiynyl, thianthrenyl,phenathridinyl, phenathrolinyl, phthalazinyl, pteridinyl, purinyl,puteridinyl, pyrazyl, pyrazolyl, pyridyl, pyridinyl, pyridazinyl,pyrazinyl, pyrimidinyl, pyrimidyl, pyrrolyl, quinazolinyl, quinolinyl,quinoxalinyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl,(1,2,3)- and (1,2,4)-triazolyl and the like, and their oxides whereappropriate, such as for example pyridyl-N-oxide.

As used herein, the phrase “substituted heteroaryl” refers to aheteroaryl group, as defined herein, in which one or more (up to aboutfive, preferably up to about three) hydrogen atoms is replaced by asubstituent independently selected from the group defined herein.

As used herein, the phrase “leaving group” refers to a group with themeaning conventionally associated with it in synthetic organicchemistry, i.e., an atom or group displaceable under substitutionreaction conditions. Examples of leaving groups include, but are notlimited to, halogen, alkane- or arylenesulfonyloxy, such asmethanesulfonyloxy, ethanesulfonyloxy, thiomethyl, benzenesulfonyloxy,tosyloxy, and thienyloxy, dihalophosphinoyloxy, optionally substitutedbenzyloxy, isopropyloxy, acyloxy, and the like. In some embodiments, aleaving group can be HC(O)—COOH or RC(O)—COOH, wherein R is a C₁-C₆alkyl or substituted C₁-C₆ alkyl.

The compounds of the invention as described herein may be synthesizedusing standard synthetic techniques known to those of skill in the artor using methods known in the art in combination with methods describedherein. The starting materials used for the synthesis of the compoundsof the invention as described herein, can be obtained from commercialsources, such as Aldrich Chemical Co. (Milwaukee, Wis.), Sigma ChemicalCo. (St. Louis, Mo.), or the starting materials can be synthesized. Thecompounds described herein, and other related compounds having differentsubstituents can be synthesized using techniques and materials known tothose of skill in the art, such as described, for example, in March,Advanced Organic Chemistry 4, Ed. (1992) John Wiley & Sons, New York,N.Y.; Carey and Sundberg, Advanced Organic Chemistry 4^(th) Ed., Vols. A(2000) and B (2001) Plenum Press, New York, N.Y. and Greene and Wuts,Protective Groups in Organic Synthesis, 3rd Ed. (1999) John Wiley &Sons, New York, N.Y., (all of which are incorporated by reference intheir entirety). General methods for the preparation of compounds asdisclosed herein may be derived from known reactions in the field, andthe reactions may be modified by the use of appropriate reagents andconditions, as would be recognized by the skilled person, for theintroduction of the various moieties found in the formulae as providedherein. For example, the compounds described herein can be modifiedusing various electrophiles or nucleophiles to form new functionalgroups or substituents.

The terms “food” or “food product” refer to a plant or plant parts.

The term “plant(s)” and “plant parts” include, but not limited to, planttissues, such as leaves, calli, stems, roots, flowers, fruits,vegetables, pollen, and seeds. A class of plants that may be used in thepresent invention is generally as broad as the class of higher and lowerplants including, but not limited to, dicotyledonous plants,monocotyledonous plants, and plant crops, including, but not limited to,vegetable crops, fruit crops, ornamental crops, and meats.

“Vegetable crops” include, but are not limited to, asparagus, beet(e.g., sugar beet and fodder beet), beans, broccoli, cabbage, carrot,cassava, cauliflower, celery, cucumber, eggplant, garlic, gherkin, leafygreens (lettuce, kale, spinach, and other leafy greens), leek, lentils,mushroom, onion, peas, pepper (e.g., sweet peppers, bell peppers, andhot peppers), potato, pumpkin, sweet potato, snap bean, squash, tomato,and turnip.

“Fruit crops” include, but are not limited to, apple, avocado, banana,soft fruits, such as, strawberry, blueberry, raspberry, blackberry,cranberry, currents and other types of soft fruit berries, carambola,cherry, citrus (e.g., oranges, lemon, lime, mandarin, grapefruit, andother citrus), coconut, fig, grapes, guava, kiwifruit, mango, nectarine,melons (including cantaloupe, muskmelon, watermelon, and other melons),olive, papaya, passionfruit, peach, pear, persimmon, pineapple, plum,and pomegranate. More specifically, horticultural crops of the presentdisclosure include, but are not limited to, soft fruits (e.g., grape,apple, pear, and persimmon) and berries (e.g., strawberries,blackberries, blueberries, and raspberries).

“Ornamental crops” include, but are not limited to, baby's breath,carnation, dahlia, daffodil, geranium, gerbera, lily, orchid, peony,Queen Anne's lace, rose, snapdragon, or other cut-flowers or ornamentalflowers, potted flowers, flower bulbs, shrub, and deciduous orconiferous tree.

“Meat” or “Meats” include, but are not limited to beef, bison, chicken,deer, goat, turkey, pork, sheep, fish, shellfish, mollusks, or dry-curedmeat products.

The term “subliming” refers the ability of a chemical, compound, orcomposition or other solid substance to evaporate or to disperse intovapor or gas when heated. Often the substance can transition back to asolid upon cooling.

The term “vaporizing” refers to transitioning or converting into vapor.

The term “volatile” or “volatilizes” refers to the ability of achemical, compound, or composition or other substance to evaporate or todisperse into vapor or gas.

Compounds and Components of the Present Methods

The methods of the present disclosure are directed to treating foodpackaging materials or containers with one or more volatileantimicrobial compounds.

More specifically, the methods described herein provide for coatingbenzoxaborole compounds on surfaces of food packaging materials orcontainers in order to delay or inhibit microorganism growth and fooddecay. Accordingly, the methods of the present disclosure comprise,consist of, or consist essentially of benzoxaborole compounds.

Exemplary embodiments of the compounds of the present disclosure aredescribed herein. In some embodiments, the antimicrobial compound,sometimes called a benzoxabroole, comprises Compounds A, B, or C, whichmay encompass diastereomers and enantiomers of the illustrativecompounds. Enantiomers are defined as one of a pair of molecularentities which are mirror images of each other and non-superimposable.Diastereomers or diastereoisomers are defined as stereoisomers otherthan enantiomers. Diastereomers or diastereoisomers are stereoisomersnot related as mirror images. Diastereoisomers are characterized bydifferences in physical properties.

One exemplary embodiment of a benzoxaborole compound of the presentmethod is Compound A:

or an analog or derivative thereof.

An additional illustrative embodiment of a benzoxaborole compound of thepresent method is Compound B:

or an analog or derivative thereof.

Another exemplary embodiment of a benzoxaborole compound of the presentmethod is Compound C, which is a salt version of Compounds A and/or B:

or an analog or derivative thereof.

In some other exemplary embodiments, the benzoxaborole, sometimes calleda volatile antimicrobial compound, of the invention has a structure offormula (I), (II), or (III):

-   -   wherein q1 and q2 are independently 1, 2, or 3;    -   q3=0, 1, 2, 3, or 4;    -   M is hydrogen, halogen, —OCH₃, or —CH₂—O—CH₂—O—CH₃;    -   M¹ is halogen, —CH₂OH, or —OCH₃;    -   X is O, S, or NR^(1c), wherein R^(1c) is hydrogen, substituted        alkyl, or unsubstituted alkyl;    -   R¹, R^(1a), R^(1b), R², and R⁵ are independently hydrogen, OH,        NH₂, SH, CN, NO₂, SO₂, OSO₂OH, OSO₂NH₂, substituted or        unsubstituted cycloalkyl, substituted or unsubstituted        heterocycloalkyl, substituted or unsubstituted aryl, or        substituted or unsubstituted heteroaryl;    -   R* is substituted or unsubstituted aryl, substituted or        unsubstituted arylalkyl, substituted or unsubstituted        heteroaryl, substituted or unsubstituted heteroarylalkyl, or        substituted or unsubstituted vinyl;    -   with a proviso that when M is F, R* is not a member selected        from:

-   -   and with a proviso that when M is Cl, R* is not a member        selected from:

-   -   and with a proviso that when M is hydrogen, R* is not a member        selected from:

-   -   wherein s=1 or 2; and R³ and R⁴ are independently methyl or        ethyl;    -   and with a provision that when M is OCH₃, R* is not a member        selected from:

-   -   and with a provision that when M¹ is F, R* is not a member        selected from:

-   -   and pharmaceutically acceptable salts thereof.    -   In one embodiment, the R* has a structure selected from:

wherein X is a member selected from CH═CH, N═CH, NR¹⁴, O and S;

wherein R¹⁴ is a member selected from H, substituted or unsubstitutedalkyl, substituted or unsubstituted aryl and substituted orunsubstituted arylalkyl;

Y is a member selected from CH and N;

-   -   R¹⁷ and R¹⁸ are members independently selected from H,        substituted or unsubstituted alkyl, substituted or unsubstituted        aryl, substituted or unsubstituted arylalkyl, (CH₂)_(v)OH,        (CH₂)_(w)NR¹⁵R¹⁶, CO₂H, CO₂-alkyl, CONH₂, S-alkyl, S-aryl,        SO-alkyl, SO-aryl, SO₂-alkyl, SO₂-aryl, SO₂H, SCF₂, CN, halogen,        CF₃ and NO₂;    -   wherein R¹⁵ and R¹⁶ are members independently selected from        hydrogen, substituted or unsubstituted alkyl and substituted or        unsubstituted alkanoyl;    -   v=1, 2, or 3; and    -   w=0, 1, 2, or 3.

In another embodiment, the R* has the following structure:

wherein R¹⁷, R¹⁸, R¹⁹, R²⁰, and R²¹ are independently selected from H,substituted or unsubstituted alkyl, substituted or unsubstituted aryl,substituted or unsubstituted arylalkyl, substituted or unsubstitutedalkyloxy, substituted or unsubstituted aryloxy, substituted orunsubstituted oxazolidin-2-yl, (CH₂)_(t)OH, CO₂H, CO₂-alkyl, CONH₂,CONH-alkyl, CON(alkyl)₂, OH, SH, S-alkyl, S-aryl, SO-alkyl, SO-aryl,SO₂-alkyl, SO₂-aryl, SO₂H, SCF₃, CN, halogen, CF₃, NO₂,(CH₂)_(u)NR²²R²³, SO₂NH₂, OCH₂CH₂NH₂, OCH₂CH₂NH-alkyl andOCH₂CH₂N(alkyl)₂;

wherein t=1, 2 or 3;

u=0, 1, or 2;

R²² and R²³ are independently selected from H, substituted orunsubstituted alkyl, and substituted or unsubstituted alkanoyl.

In another embodiment, the R* has the following structure:

wherein R¹⁷, R¹⁸, R¹⁹, R²⁰, and R²¹ are independently selected from H,substituted or unsubstituted alkyl, substituted or unsubstituted aryl,substituted or unsubstituted arylalkyl, substituted or unsubstitutedalkyloxy, substituted or unsubstituted aryloxy, substituted orunsubstituted oxazolidin-2-yl, (CH₂)_(t)OH, CO₂H, CO₂-alkyl, CONH₂,CONH-alkyl, CON(alkyl)₂, OH, SH, S-alkyl, S-aryl, SO-alkyl, SO-aryl,SO₂-alkyl, SO₂-aryl, SO₂H, SCF₃, CN, halogen, CF₃, NO₂,(CH₂)_(u)NR²²R²³, SO₂NH₂, OCH₂CH₂NH₂, OCH₂CH₂NH-alkyl andOCH₂CH₂N(alkyl)₂;

wherein t=1, 2 or 3;

u=0, 1, or 2;

R²² and R²³ are independently selected from H, substituted orunsubstituted alkyl, and substituted or unsubstituted alkanoyl;

R²⁴ and R²⁵ are independently selected from H, substituted orunsubstituted alkyl, substituted or unsubstituted aryl, substituted orunsubstituted arylalkyl, substituted or unsubstituted alkyloxy,substituted or unsubstituted aryloxy, substituted or unsubstitutedoxazolidin-2-yl, (CH₂), OH, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl,CON(alkyl)₂, OH, SH, S-alkyl, S-aryl, SO-alkyl, SO-aryl, SO₂-alkyl,SO₂-aryl, SO₃H, SCF₃, CN, halogen, CF₃, NO₂, (CH₂)_(u)NR²²R²³, SO₂NH₂,OCH₂CH₂NH₂, OCH₂CH₂NH-alkyl and OCH₂CH₂N(alkyl)₂;

Z=1, 2, 3, 4, 5, or 6.

Additional antimicrobial compounds are also disclosed previously in U.S.Pat. No. 8,106,031, and International Patent Application WO2007/131072A2, the contents of which are hereby incorporated byreference in their entireties.

In some embodiments, the volatile antimicrobial compound of theinvention has the structure of formula (IV):

wherein A and D together with the carbon atoms to which they areattached form a 5-, 6-, or 7-membered fused ring which may besubstituted by C₁-C₆-alkyl, C₁-C₆-alkoxy, hydroxy, halogen, nitro,nitrile, amino, amino substituted by one or more C₁-C₆-alkyl groups,carboxy, acyl, aryloxy, carbonamido, carbonamido substituted byC₁-C₆-alkyl, sulfonamido or trifluoromethyl or the fused ring may linktwo oxaborole rings:

X is a group —CR⁷R⁸ wherein R⁷ and R⁸ are each independently hydrogen,C₁-C₆-alkyl, nitrile, nitro, aryl, arylalkyl or R⁷ and R⁸ together withthe carbon atom to which they are attached form an alicyclic ring; and

R⁶ is hydrogen, C₁-C₁₈-alkyl, C₁-C₁₈-alkyl substituted by C₁-C₆-alkoxy.C₁-C₆-alkylthio, hydroxy, amino, amino substituted by C₁-C₁₈-alkyl,carboxy, aryl, aryloxy, carbonamido, carbonamido substituted byC₁-C₆-alkyl, aryl or arylalkyl, arylalkyl, aryl, heteroaryl, cycloalkyl.C₁-C₁₈-alkyleneamino, C₁-C₁₈-alkyleneamino substituted by phenyl.C₁-C₆-alkoxy or C₁-C₆-alkylthio, carbonyl alkylencamino or a radical offormula (V):

wherein A, D and X are as defined herein before except forboronophthalide;

and pharmaceutically acceptable salts thereof.

In one embodiment, the volatile antimicrobial compound of the inventionhas the structure of formula (IX):

wherein A, D, and X are defined as above;

Y is a divalent alkylene linking group containing up to 18 carbon atomsor a divalent alkylene linking group containing up to 18 carbon atomswhich is substituted by phenyl, C₁-C₆ alkoxy, C₁-C₆-alkylthio; carbonylalkylene amino; and

R³ and R⁴ are each, independently, hydrogen, C₁-C₁₈-alkyl or phenyl orR³ together with Y or part of Y forms a 5-, 6- or 7-membered ringcontaining the nitrogen atom.

In another embodiment, the volatile antimicrobial compound of theinvention has the structure of formula (X):

wherein A, D, and X are defined as above;

n is 1, 2, or 3;

R³ is hydrogen, C₁-C₁₈-alkyl or phenyl; and

R⁵ and R⁶ are each, independently, hydrogen, alkyl containing up to atotal of 16 carbon atoms or phenyl.

Additional antimicrobial compounds are also disclosed previously in U.S.Pat. No. 5,880,188, the content of which is hereby incorporated byreference in its entirety.

In another aspect, the volatile antimicrobial compound of the inventionhas the structure of formula (VI):

wherein each R is independently hydrogen, alkyl, alkene, alkyne,haloalkyl, haloalkene, haloalkyne, alkoxy, alkeneoxy, haloalkoxy, aryl,heteroaryl, arylalkyl, arylalkene, arylalkyne, heteroarylalkyl,heteroarylalkene, heteroarylalkyne, halogen, hydroxyl, nitrile, amine,ester, carboxylic acid, ketone, alcohol, sulfide, sulfoxide, sulfone,sulfoximine, sulfilimine, sulfonamide, sulfate, sulfonate, nitroalkyl,amide, oxime, imine, hydroxylamine, hydrazine, hydrazone, carbamate,thiocarbamate, urea, thiourea, carbonate, aryloxy, or heteroaryloxy;

n=1, 2, 3, or 4;

B is boron;

X═(CR₂)_(m) where m=1, 2, 3, or 4;

Y is alkyl, alkene, alkyne, haloalkyl, haloalkene, haloalkyne, alkoxy,alkeneoxy, haloalkoxy, aryl, heteroaryl, arylalkyl, arylalkene,arylalkyne, heteroarylalkyl, heteroarylalkene, heteroarylalkyne,hydroxyl, nitrile, amine, ester, carboxylic acid, ketone, alcohol,sulfide, sulfoxide, sulfone, sulfoximine, sulfilimine, sulfonamide,sulfate, sulfonate, nitroalkyl, amide, oxime, imine, hydroxylamine,hydrazine, hydrazone, carbamate, thiocarbamate, urea, thiourea,carbonate, aryloxy, or heteroaryloxy;

with a proviso that R is not aryloxy or heteroaryloxy when Y ishydroxyl;

and pharmaceutically acceptable salts thereof.

In one embodiment, the volatile antimicrobial compound has a structureof formula (VII):

wherein W═(CH₂)_(q) where q is 1, 2, or 3.

In another embodiment, the volatile antimicrobial compound has astructure of

In another embodiment, the volatile antimicrobial compound of theinvention has the structure of formula (VIII):

wherein R^(a) is CN, C(O)NR⁹R¹⁰, or C(O)OR¹¹ wherein R¹¹ is hydrogen,substituted alkyl, or unsubstituted alkyl,

X is N, CH and CR^(b);

R^(b) is halogen, substituted or unsubstituted alkyl, C(O)R¹², C(O)OR¹²,OR¹², NR¹²R¹³, wherein R⁹, R¹⁰, R¹², and R¹³ are independently hydrogen,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl;

with a proviso that R⁹ and R¹⁰, together with the atoms to which theyare attached, are optionally combined to form a 4- to 8-memberedsubstituted or unsubstituted heterocycloalkyl ring;

and with a proviso that R¹² and R¹³, together with the atoms to whichthey are attached, are optionally combined to form a 4- to 8-memberedsubstituted or unsubstituted heterocycloalkyl ring;

and pharmaceutically acceptable salts thereof.

In one embodiment, the volatile antimicrobial compound of the inventionhas the structure of formula (XI):

In another embodiment, the volatile antimicrobial compound of theinvention is selected from:

In another embodiment, the volatile antimicrobial compound of theinvention is selected from:

In another embodiment, the volatile antimicrobial compound of theinvention is selected from:

In one embodiment, the volatile antimicrobial compound of the inventionhas the structure of formula (XII):

In another embodiment, the volatile antimicrobial compound of theinvention is selected from:

wherein R³ is hydrogen, substituted or unsubstituted alkyl, orsubstituted or unsubstituted heteroalkyl.

In another embodiment, the volatile antimicrobial compound of theinvention is selected from:

wherein R³ is hydrogen, substituted or unsubstituted alkyl, orsubstituted or unsubstituted heteroalkyl.

In another embodiment, the volatile antimicrobial compound of theinvention is selected from:

wherein R³ is hydrogen, substituted or unsubstituted alkyl, orsubstituted or unsubstituted heteroalkyl.

In one embodiment, the volatile antimicrobial compound of the inventionhas the structure of formula (XIII):

wherein each of R¹ and R² is independently hydrogen, substituted orunsubstituted alkyl, or substituted or unsubstituted heteroalkyl.

In another embodiment, the volatile antimicrobial compound of theinvention is selected from:

In another embodiment, the volatile antimicrobial compound of theinvention is selected from:

wherein each of R¹ and R² is independently hydrogen, substituted orunsubstituted alkyl, or substituted or unsubstituted heteroalkyl.

In another embodiment, the volatile antimicrobial compound of theinvention is selected from:

wherein each of R¹ and R² is independently hydrogen, substituted orunsubstituted alkyl, or substituted or unsubstituted heteroalkyl.

In one embodiment, R^(b) is selected from fluorine and chlorine. Inanother embodiment, R^(b) is selected from OR²⁶ and NR²⁷R²⁸. In anotherembodiment when R^(b) is OR²⁶, R²⁶ is selected from H, substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, and substituted orunsubstituted heteroaryl. In another embodiment when R^(b) is OR²⁶, R²⁶is selected from H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl and substituted or unsubstituted cycloalkyl.In another embodiment when R^(b) is OR²⁶, R²⁶ is unsubstituted C₁-C₆alkyl. In another embodiment when R^(b) is OR²⁶, R²⁶ is unsubstitutedcycloalkyl. In another embodiment when R^(b) is OR²⁶, R²⁶ is alkyl,substituted with a member selected from substituted or unsubstitutedC₁-C₆ alkoxy. In another embodiment when R^(b) is OR²⁶, R²⁶ is alkyl,substituted with at least one halogen. In another embodiment when R^(b)is OR²⁶, R²⁶ is alkyl, substituted with at least one oxo moiety.

In another embodiment when R^(b) is OR²⁶, R²⁶ is a member selected from—CH₃, —CH₂CH₃, —(CH₂)₂CH₃, —CH(CH₃)₂, —CH₂CF₃, —CH₂CHF₂, —CH₂CH₂(OH),—CH₂CH₂(OCH₃), —CH₂CH₂(OC(CH₃)₂), —C(O)CH₃, —CH₂CH₂OC(O)CH₃,—CH₂C(O)OCH₂CH₃, —CH₂C(O)OC(CH₃)₃, —(CH₂)₃C(O)CH₃, —CH₂C(O)OC(CH₃)₃,cyclopentyl, cyclohexyl,

In another embodiment when R^(b) is NR²⁷R²⁸, R²⁷ and R²⁸ are membersindependently selected from H, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, and substituted or unsubstituted heteroaryl. Inanother embodiment when R^(b) is NR²⁷R²⁸, R²⁷ is H or unsubstitutedalkyl; and R²⁸ is unsubstituted alkyl or alkyl substituted with a memberselected from hydroxyl, phenyl, unsubstituted alkoxy and alkoxysubstituted with a phenyl. In a further embodiment when R^(b) isNR²⁷R²⁸, R²⁷ is H or CH₃.

In another embodiment when R^(b) is NR²⁷R²⁸, R²⁷ and R²⁸ areindependently selected from substituted or unsubstituted alkyl. Inanother embodiment when R^(b) is NR²⁷R²⁸, R²⁷ is unsubstituted alkyl;and R²⁸ is substituted or unsubstituted alkyl. In another embodimentwhen R^(b) is NR²⁷R²⁸, R²⁷ is unsubstituted alkyl; and R²⁸ is alkyl,substituted with a member selected from substituted or unsubstitutedalkoxy and hydroxyl. In another embodiment when R^(b) is NR²⁷R²⁸, R²⁷ isunsubstituted alkyl; and R²⁸ is alkyl, substituted with unsubstitutedalkoxy. In another embodiment when R^(b) is NR²⁷R²⁸, R²⁷ isunsubstituted alkyl; and R²⁸ is alkyl, substituted with alkoxy,substituted with phenyl. In another embodiment when R^(b) is NR²⁷R²⁸,R²⁷ is unsubstituted alkyl; and R²⁸ is alkyl, substituted withunsubstituted alkoxy. In another embodiment when R^(b) is NR²⁷R²⁸, R²⁷and R²⁸ together with the nitrogen to which they are attached, arecombined to form a 4- to 8-membered substituted or unsubstitutedheterocycloalkyl ring. In another embodiment when R^(b) is NR²⁷R²⁸, R²⁷and R²⁸ together with the nitrogen to which they are attached, arecombined to form a 5- or 6-membered substituted or unsubstitutedheterocycloalkyl ring.

In another embodiment, R^(b) is selected from N(CH₃)₂,N(CH₃)(CH₂CH₂(OCH₃)), N(CH₃)(CH₂CH₂OH), NH₂, NHCH₃, NH(CH₂CH₂(OCH₃)),NH(CH₂CH₂(OCH₂Ph), NH(CH₂Ph), NH(C(CH₃)₃) and NH(CH₂CH₂OH). In anotherembodiment, R^(b) is selected from

Additional antimicrobial compounds are also disclosed previously in U.S.Pat. No. 8,039,450, and patent application publication US 2009/0291917,the contents of which are hereby incorporated by reference in theirentireties.

In one aspect, the volatile antimicrobial compound of the invention hasthe structure of formula (A):R^(A)-L^(A)-G-L^(B)-R^(B)  (A),whereineach of R^(A) and R^(B) is independently a radical comprising anoxaborole moiety;each of L^(A) and L^(B) is independently —O— or

each of R and R′ is independently hydrogen, unsubstituted or substitutedC₁₋₁₈-alkyl, arylalkyl, aryl, or heterocyclic moiety; andG is a substituted or unsubstituted C₁₋₁₈-alkylene, arylalkylene,arylene, or heterocyclic moiety; and pharmaceutically acceptable saltsthereof.

In one embodiment, the volatile compound is an antimicrobial compound.In another embodiment, the volatile compound has use against pathogensaffecting meats, plants, or plant parts, comprising contacting themeats, plants, or plant parts. In another embodiment, the-L^(A)-G-L^(B)- portion of formula (A) is derived from a diol or diaminecompound. In a further embodiment, the diol compound is selected fromthe group consisting of 1,2-ethylene glycol; 1,2-propylene glycol;1,3-propylene glycol; 1,1,2,2-tetramethyl-1,2-ethylene glycol;2,2-dimethyl-1,3-propylene glycol; 1,6-hexanediol; 1,10-decanediol; andcombinations thereof. In another embodiment, the diamine compound is1,2-ethylene diamine; 1,3-propylene diamine; or combinations thereof. Inanother embodiment, L^(A) and L^(B) are identical. In anotherembodiment, L^(A) and L^(B) are different. In another embodiment, eachof L^(A) and L^(B) is independently —O— or —NH—. In another embodiment,L^(A) and L^(B) are identical. In another embodiment, L^(A) and L^(B)are different.

In another embodiment, the -L^(A)-G-L^(B)- portion of formula (A)comprises asymmetrical functional groups (i.e., asymmetrical bridges).In a further embodiment, the -L^(A)-G-L^(B)- portion of formula (A)comprises one hydroxyl group and one amine group. In a furtherembodiment, the -L^(A)-G-L^(B)- portion of formula (A) comprises anamino alcohol. In another embodiment, G is a substituted orunsubstituted C₁₋₈-alkylene. In a further embodiment, G is a substitutedor unsubstituted C₁₋₄-alkylene. In a further embodiment, G is selectedfrom —CH₂—, —CH₂—CH₂—, and —CH₂—CH₂—CH₂—.

In another embodiment, each of R^(A) and R^(B) is independently derivedfrom the group consisting of5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole;5-chloro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole;1,3-dihydro-1-hydroxy-2,1-benzoxaborole; and combinations thereof. Inanother embodiment, R^(A) and R^(B) are identical. In anotherembodiment, R^(A) and R^(B) are different.

In another embodiment, at least one of R^(A) and R^(B) is selected fromformula (B), (C), or (D):

-   -   wherein q1 and q2 are independently 1, 2, or 3;    -   q3=0, 1, 2, 3, or 4;    -   B is boron;    -   M is hydrogen, halogen, —OCH₃, or —CH₂—O—CH₂—O—CH₃;    -   M¹ is halogen, —CH₂OH, or —OCH₃;    -   X is O, S, or NR^(1c), wherein R^(1c) is hydrogen, substituted        alkyl, or unsubstituted alkyl;    -   R¹, R^(1a), R^(1b), R², and R⁵ are independently hydrogen, OH,        NH₂, SH, CN, NO₂, SO₂, OSO₂OH, OSO₂NH₂, substituted or        unsubstituted cycloalkyl, substituted or unsubstituted        heterocycloalkyl, substituted or unsubstituted aryl, or        substituted or unsubstituted heteroaryl;

and pharmaceutically acceptable salts thereof.

Additional oxaborole moieties are also disclosed previously in U.S. Pat.No. 8,106,031, and International Patent Application WO 2007/131072A2,the contents of which are hereby incorporated by reference in theirentireties.

In another embodiment, at least one of R^(A) and R^(B) has a structureof formula (F):

wherein A and D together with the carbon atoms to which they areattached form a 5, 6, or 7-membered fused ring which may be substitutedby C₁₋₆-alkyl, C₁₋₆-alkoxy, hydroxy, halogen, nitro, nitrile, amino,amino substituted by one or more C₁₋₆-alkyl groups, carboxy, acyl,aryloxy, carbonamido, carbonamido substituted by C₁₋₆-alkyl,sulphonamido or trifluoromethyl or the fused ring may link two oxaborolerings; B is boron;

X¹ is a group —CR⁷R⁸ wherein R⁷ and R⁸ are each independently hydrogen,C₁₋₆-alkyl, nitrile, nitro, aryl, aralkyl or R⁷ and R⁸ together with thecarbon atom to which they are attached form an alicyclic ring; and

and pharmaceutically acceptable salts thereof.

Additional oxaborole moieties are also disclosed previously in U.S. Pat.No. 5,880,188, the content of which is hereby incorporated by referencein its entirety.

In another embodiment, at least one of R^(A) and R^(B) is selected fromformula (E) or (G):

wherein each R⁶ is independently hydrogen, alkyl, alkene, alkyne,haloalkyl, haloalkene, haloalkyne, alkoxy, alkeneoxy, haloalkoxy, aryl,heteroaryl, arylalkyl, arylalkene, arylalkyne, heteroarylalkyl,heteroarylalkene, heteroarylalkyne, halogen, hydroxyl, nitrile, amine,ester, carboxylic acid, ketone, alcohol, sulfide, sulfoxide, sulfone,sulfoximine, sulfilimine, sulfonamide, sulfate, sulfonate, nitroalkyl,amide, oxime, imine, hydroxylamine, hydrazine, hydrazone, carbamate,thiocarbamate, urea, thiourea, carbonate, aryloxy, or heteroaryloxy;

n=1, 2, 3, or 4;

B is boron;

X²═(CR⁶ ₂)_(m) where m=1, 2, 3, or 4; or

wherein R⁹ is CN, C(O)NR¹¹R¹², or C(O)OR³ wherein R³ is hydrogen,substituted alkyl, or unsubstituted alkyl,

X³ is N, CH and CR¹⁰;

R¹⁰ is halogen, substituted or unsubstituted alkyl, C(O)R¹⁴, C(O)OR¹⁴,OR¹⁴, NR¹⁴R¹⁵, wherein each of R¹¹, R¹², R¹⁴, and R¹⁵ is independentlyhydrogen, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl;

and pharmaceutically acceptable salts thereof.

In a further embodiment when at least one of R^(A) and R^(B) has astructure of formula (G), R⁹ is CN and R¹⁰ is R^(b).

In another embodiment, at least one of R^(A) and R^(B) has a structureselected from:

In another embodiment, at least one of R^(A) and R^(B) has a structureselected from:

In another embodiment, at least one of R^(A) and R^(B) has a structureselected from:

In another embodiment when at least one of R^(A) and R^(B) has astructure of formula (G), R⁹ is —COOR³ and R¹⁰ is R^(b).

In another embodiment, at least one of R^(A) and R^(B) has a structureselected from:

In another embodiment, at least one of R^(A) and R^(B) has a structureselected from:

In another embodiment, at least one of R^(A) and R^(B) has a structureselected from:

In another embodiment when at least one of R^(A) and R^(B) has astructure of formula (G), R⁹ is —CONR¹R² and R¹⁰ is R^(b).

In another embodiment, each of R^(A) and R^(B) is independently selectedfrom formula (B), (C), (D), (E), (F), or (G).

In another embodiment, the volatile compound of the invention isselected from:

In another embodiment, the volatile compound of the invention isselected from:

In another embodiment, the volatile compound of the invention isselected from:

In one embodiment, R^(b) is selected from fluorine and chlorine. Inanother embodiment, R^(b) is selected from OR²⁰ and NR²¹R²². In anotherembodiment when R^(b) is OR²⁰, R²⁰ is selected from H, substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, and substituted orunsubstituted heteroaryl. In another embodiment when R^(b) is OR²⁰, R²⁰is selected from H, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl and substituted or unsubstituted cycloalkyl.In another embodiment when R^(b) is OR²⁰, R²⁰ is unsubstituted C₁₋₆alkyl. In another embodiment when R^(b) is OR²⁰, R²⁰ is unsubstitutedcycloalkyl. In another embodiment when R^(b) is OR²⁰, R²⁰ is alkyl,substituted with a member selected from substituted or unsubstitutedC₁₋₆ alkoxy. In another embodiment when R^(b) is OR²⁰, R²⁰ is alkyl,substituted with at least one halogen. In another embodiment when R^(b)OR²⁰, R²⁰ is alkyl, substituted with at least one oxo moiety.

In another embodiment when R^(b) is OR²⁰, R²⁰ is a member selected from—CH₃, —CH₂CH₃, —(CH₂)₂CH₃, —CH(CH₃)₂, —CH₂CF₃, —CH₂CHF₂, —CH₂CH₂(OH),—CH₂CH₂(OCH₃), —CH₂CH₂(OC(CH₃)₂), —C(O)CH₃, —CH₂CH₂OC(O)CH₃,—CH₂C(O)OCH₂CH₃, —CH₂C(O)OC(CH₃)₃, —(CH₂)₃C(O)CH₃, —CH₂C(O)OC(CH₃)₃,cyclopentyl, cyclohexyl,

In another embodiment when R^(b) is NR²¹R²², R²¹ and R²² are membersindependently selected from H, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, and substituted or unsubstituted heteroaryl. Inanother embodiment when R^(b) is NR²¹R²², R²¹ is H or unsubstitutedalkyl; and R²² is unsubstituted alkyl or alkyl substituted with a memberselected from hydroxyl, phenyl, unsubstituted alkoxy and alkoxysubstituted with a phenyl. In a further embodiment when R^(b) isNR²¹R²², R²¹ is H or CH₃.

In another embodiment when R^(b) is NR²¹R²², R²¹ and R²² areindependently selected from substituted or unsubstituted alkyl. Inanother embodiment when R^(b) is NR²¹R²², R²¹ is unsubstituted alkyl;and R²² is substituted or unsubstituted alkyl. In another embodimentwhen R^(b) is NR²¹R²², R²¹ is unsubstituted alkyl; and R²² is alkyl,substituted with a member selected from substituted or unsubstitutedalkoxy and hydroxyl. In another embodiment when R^(b) is NR²¹R²², R²¹ isunsubstituted alkyl; and R²² is alkyl, substituted with unsubstitutedalkoxy. In another embodiment when R^(b) is NR²¹R²², R²¹ isunsubstituted alkyl; and R²² is alkyl, substituted with alkoxy,substituted with phenyl. In another embodiment when R^(b) is NR²¹R²²,R²¹ is unsubstituted alkyl; and R²² is alkyl, substituted withunsubstituted alkoxy. In another embodiment when R^(b) is NR²¹R²², R²¹and R²² together with the nitrogen to which they are attached, arecombined to form a 4- to 8-membered substituted or unsubstitutedheterocycloalkyl ring. In another embodiment when R^(b) is NR²¹R²², R²¹and R²² together with the nitrogen to which they are attached, arecombined to form a 5- or 6-membered substituted or unsubstitutedheterocycloalkyl ring.

In another embodiment, R^(b) is selected from N(CH₃)₂,N(CH₃)(CH₂CH₂(OCH₃)), N(CH₃)(CH₂CH₂OH), NH₂, NHCH₃, NH(CH₂CH₂(OCH₃)),NH(CH₂CH₂(OCH₂Ph), NH(CH₂Ph), NH(C(CH₃)₃) and NH(CH₂CH₂OH). In anotherembodiment, R^(b) is selected from

Additional oxaborole moieties are also disclosed previously in U.S. Pat.No. 8,039,450, and patent application publication US 2009/0291917, thecontents of which are hereby incorporated by reference in theirentireties.

In another embodiment, the compound provided has a structure of formula(A1) or (A2):

wherein each of A¹, A², D¹, and D² is independently hydrogen,substituted or unsubstituted C₁₋₁₈-alkyl, arylalkyl, aryl, orheterocyclic; or A¹ and D¹, or A² and D² together form a 5, 6, or7-membered fused ring which is substituted or unsubstituted;

each of R¹³, R¹⁶, R¹⁷, R¹⁸, and R¹⁹ is independently hydrogen,substituted or unsubstituted C₁₋₆-alkyl, nitrile, nitro, aryl or arylalkyl; or R¹⁶ and R¹⁷, or R¹⁸ and R¹⁹ together form an alicyclic ringwhich is substituted or unsubstituted;

B is boron; and

G is a substituted or unsubstituted C₁₋₁₈-alkylene, arylalkylene,arylene, or heterocyclic moiety.

In another embodiment, each of R^(A) and R^(B) is independently

wherein X²═(CR⁶ ₂)_(m) and m=1, 2, 3, or 4.

In another embodiment, each of R^(A) and R^(B) is independently

In another embodiment, the compound provided has the structure of

Additional oxaborole moieties are also disclosed previously in U.S. Pat.No. 5,880,188, the content of which is hereby incorporated by referencein its entirety.

In another aspect, the antimicrobial compound, sometimes called abenzoxaborole, is a compound having a structure of formula (AX):

wherein A and D together with the carbon atoms to which they areattached form a 5-, 6-, or 7-membered fused ring which may besubstituted by C₁-C₆-alkyl, C₁-C₆-alkoxy, hydroxy, halogen, nitro,nitrile, amino, amino substituted by one or more C₁-C₆-alkyl groups,carboxy, acyl, aryloxy, carbonamido, carbonamido substituted byC₁-C₆-alkyl, sulfonamido or trifluoromethyl or the fused ring may linktwo oxaborole rings; B is boron;

R¹ and R² are each independently halogen or nitrile;

X¹ is a group —(CR³R⁴)_(p) wherein R³ and R⁴ are each independentlyhydrogen, C₁-C₆-alkyl, nitrile, nitro, aryl, arylalkyl or R³ and R⁴together with the carbon atom to which they are attached form analicyclic ring;

p is 1, 2, 3, or 4;

M⁺ is a counterion;

and agriculturally acceptable salts thereof.

Additional disclosure and methods for making a compound of formula AXcan be found in U.S. Pat. No. 9,730,454, issued Aug. 15, 2017, thedisclosure of which is incorporated by reference in its entirety.

In one embodiment, the compound is volatile. In another embodiment, thecompound has antimicrobial activity.

In one embodiment, the compound of formula (AX) is prepared from a(precursor) compound selected from the group consisting of5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole;5-chloro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole;1,3-dihydro-1-hydroxy-2,1-benzoxaborole; and combinations thereof. Inanother embodiment, the compound of formula (A) is prepared from a(precursor) compound selected from the group consisting of5-fluorobenzo[c][1,2]oxaborol-1(3H)-ol;5-chlorobenzo[c][1,2]oxaborol-1(3H)-ol; benzo[c][1,2]oxaborol-1(3H)-ol;and combinations thereof.

In another embodiment, the compound of formula (AX) is

In a further embodiment, the compound of formula (AX) is selected fromthe group consisting of

and combinations thereof. In another embodiment, the compound of formula(A) is selected from the group consisting of

and combination thereof. In another embodiment, the compound of formula(AX) is

Additional oxaborole compounds useful for preparing compounds of formula(AX) are also disclosed in U.S. Pat. No. 5,880,188, the content of whichis hereby incorporated by reference in its entirety. In another aspect,provided is a mixture or composition comprising the compound of formula(AX).

In another aspect, provided is a method of using a compound againstpathogens affecting meats, plants, or plant parts, comprising contactingthe meats, plants, or plant parts with an effective amount of thecompound having a structure of formula (A):

wherein A and D together with the carbon atoms to which they areattached form a 5-, 6-, or 7-membered fused ring which may besubstituted by C₁-C₆-alkyl, C₁-C₆-alkoxy, hydroxy, halogen, nitro,nitrile, amino, amino substituted by one or more C₁-C₆-alkyl groups,carboxy, acyl, aryloxy, carbonamido, carbonamido substituted byC₁-C₆-alkyl, sulfonamido or trifluoromethyl or the fused ring may linktwo oxaborole rings; B is boron;

R¹ and R² are each independently halogen or nitrile;

X¹ is a group —(CR³R⁴)_(p) wherein R³ and R⁴ are each independentlyhydrogen, C₁-C₆-alkyl, nitrile, nitro, aryl, arylalkyl or R³ and R⁴together with the carbon atom to which they are attached form analicyclic ring;

p is 1, 2, 3, or 4;

and agriculturally acceptable salts thereof.

In one embodiment, the compound is volatile. In another embodiment, thecompound is a fungicide. In another embodiment, the contacting comprisesdirect contact or contact as a volatile compound, i.e., via directcontact or via volatile activity.

In a further embodiment, the contacting comprises application of aliquid formulation.

In one embodiment, the method of using a volatile compound againstpathogens affecting meats, plants, or plant parts, comprises

providing a compound of formula (AX) in gaseous form; and

contacting a meat, plant, or plant part with an effective amount of thecompound of formula (AX) in gaseous form.

In another embodiment, the method of using a volatile compound againstpathogens affecting meats, plants, or plant parts, comprises

placing a meat, plant, or plant part in a container; and

introducing into the container and in contact with the meat, plant, orplant part an effective amount of the compound of formula (A) in gaseousform.

In another embodiment, the method of using a volatile compound againstpathogens affecting meats, plants, or plant parts, comprises contactingthe meats, plants, or plant parts with an atmosphere comprising aneffective amount of the compound of formula (A) in gaseous form.

In one embodiment, the compound of formula (A) is prepared from a(precursor) compound selected from the group consisting of5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole;5-chloro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole;1,3-dihydro-1-hydroxy-2,1-benzoxaborole; and combinations thereof. Inanother embodiment, the compound of formula (A) is prepared from a(precursor) compound selected from the group consisting of5-fluorobenzo[c][1,2]oxaborol-1(3H)-ol;5-chlorobenzo[c][1,2]oxaborol-1(3H)-ol; benzo[c][1,2]oxaborol-1(3H)-ol;and combinations thereof.

In another embodiment, the compound of formula (AX) is

In a further embodiment, the compound of formula (AX) is selected fromthe group consisting of

and combinations thereof. In another embodiment, the compound of formula(A) is selected from the group consisting of

and combination thereof. In another embodiment, the compound of formula(AX) is

In some aspects, M⁺ is an alkali metal ion.

Compounds A, B, and/or C may be used individually or as a mixture orcombination. The benzoxaborole compounds may also be used in combinationwith a carrier to form a benzoxaborole treatment. The benzoxaboroletreatment provides antimicrobial protection to food, such as plants,crops, or meats, when administered, applied, or exposed to the plants,crops, or meats. While the mechanism of action of the benzoxaborolecompound is not fully understood, it is thought to proceed via blockingor inhibition of protein synthesis in microorganisms and/or blockingcytoplasmic leucyl-tRNA synthetase (LeuRS) activity, thereby preventinggrowth of microorganisms on food.

Benzoxaboroles, including Compounds A, B, and/or C, may be used in anyform, including, but not limited to, a liquid, a solid, or a gaseouscomposition. In particular, the present method provides application of abenzoxaborole compound to food product packaging materials as a spray, amist, a gel, a thermal and non-thermal fog, a dip, a drench, a vapor, agas, or via sublimation.

Carriers of the present disclosure may be combined with one or moreactive benzoxaborole compounds to form a benzoxaborole treatment.Treatment carriers of the present disclosure may comprise gases,solutions, solvents, or chemicals. For example, a liquid carrier of thepresent disclosure may comprise water, buffer, saline solution, asolvent, or solvent-based solution, etc. Illustrative liquid solventcarriers of the present disclosure include, but are not limited to,liquid carbon dioxide (CO₂), such as supercritical CO₂. Gaseous carriersof the benzoxaborole compounds may comprise nitrogen (N₂), carbondioxide (CO₂), or sulfur dioxide (SO₂).

Benzoxaborole compound treatments, such as those comprising Compounds A,B, and/or C, with or without a carrier, may be applied to packagingmaterials. More specifically, the benzoxaborole treatments of thepresent disclosure may be applied, imbedded, impregnated, or coated ontoor into one or more surfaces of a packaging material (e.g., PETclamshells or liner materials) or a device (e.g., a container or achamber), collectively and interchangeably called a “chamber.” Whencoated onto or imbedded into surfaces of packaging materials, thebenzoxaborole compound treatment volatilizes to treat food products,which ultimately preserves freshness of the food. Thus, the compoundtreatments of the present disclosure allow for the uniform treatment offood packaging materials to protect food products comprised therein.

A chamber of the present disclosure may be any container in which a foodproduct may be comprised therein for harvest, storage, and/or retailusage. For example, a chamber may be made of any material sufficient tohold food including, but not limited to, cardboard, paper, paperboard,corrugated paper, plastic (e.g., thermosets and thermoplastics), glass,polystyrene, cellulosic material, metals (e.g., aluminum, foils,laminates, tinplate, and/or steel, such as tin-free steel), or any othersemipermeable or impermeable material. Exemplary chambers of the presentdisclosure may be made of polyester, such as polycarbonate, polyethylenenaphthalate, and polyethylene terephthalate (i.e., PET or PETE). Thus,an illustrative embodiment of a chamber of the present disclosure is aPET clamshell.

The chamber of the present disclosure may be of any size to hold foodproducts within the packaging materials, such as an individual orsingulated chamber embodiment. For example, illustrative individualchambers may have a volume ranging from about 0.1 liters (L) to about 50L, from about 0.1 L to about 40 L, from about 0.1 L to about 30 L, fromabout 0.1 L to about 20 L, from about 0.1 L to about 10 L, from about0.1 L to about 5 L, from about 0.1 L to about 4 L, from about 0.1 L toabout 3 L, from about 0.1 L to about 2 L, from about 1 L to about 50 L,from about 5 L to about 40 L, from about 20 L to about 40 L, from about25 L to about 50 L, from about 30 L to about 40 L, from about 35 L toabout 40 L, and at about 0.1 L, about 0.2 L, about 0.3 L, about 0.4 L,about 0.5 L, about 1 L, about 2 L, about 10 L, about 20 L, about 30 L,about 35 L, about 40 L, and about 50 L.

An additional chamber embodiment may be capable of holding a pluralityof individual chambers. A plurality of individual chambers, such a PETclamshells, may include two or more to thousands, to many thousands totens or hundreds of thousands or millions of PET clamshells. Forexample, one box may comprise about 384 clamshells, and a chamberembodiment may comprise thousands of boxes of clamshells (e.g., fromabout 384,000 to about 3,840,000 to about 384,000,000 of clamshells).Thus, a plurality of PET clamshells is also an illustrative embodimentof the chamber of the present disclosure, which is particularly utilizedfor large-scale and/or commercial treatment methods of chambers.

A further embodiment of the chamber of the present disclosure maycomprise a liquid-absorbing material. The liquid-absorbing material maybe comprised within the chamber, such as on the internal top, bottom orside panels of the chamber. The liquid-absorbing material may becomprised on the exterior of the chamber, such as on the external top,bottom or side panels of the chamber. The liquid-absorbing material mayalso be comprised on or in one or more liners, wrapping, labels, tags,stickers, pads, or other packing components located, attached, and/oraffixed to the inside or the outside of the chamber.

The liquid-absorbing material may comprise any material that is able toabsorb and retain a liquid composition of the active compound. Forexample, illustrative embodiments of the liquid-absorbing materialinclude, but are not limited to, cotton, paper, foam, etc.

Absorption of an active ingredient (i.e., benzoxaborole) into theliquid-absorbing material enables the liquid-absorbing material to serveas a reservoir capable of releasing the benzoxaborole treatment to thefood product comprised in the chamber over a time period. Theliquid-absorbing material may provide for slow-release or quick-releaseof the benzoxaborole treatment to the food product. Thus, theliquid-absorbing material enables differential treatment of the foodproduct based on the time period required for protection of foodproducts comprised therein. For example, food products that need limitedantimicrobial protection may be packaged in a chamber comprising aquick-release liquid-absorbing material, while food products requiringan extended term of antimicrobial protection may be packaged in achamber comprising a slow-release liquid-absorbing material.

Slow-release liquid-absorbing materials include, but are not limited to,materials that enable the release of the active ingredient to the foodproduct for a time period of over 12 hours, such as from over 12 hoursto about 31 days, including from over 12 hours to about 25 days, fromover 12 hours to about 20 days, from over 12 hours to about 15 days,from over 12 hours to about 10 days, from over 12 hours to about 5 days,from over 12 hours to about 30 days, from over 12 hours to about 24days, from about 24 hours to about 30 days, from about 2 days to about28 days, from about 3 days to about 25 days, from about 4 days to about20 days, and about 5 days, about 10 days, about 15 days, about 20 days,about 25 days, about 30 days, and any number of days between 1 day to 30days.

Quick-release liquid-absorbing materials include, but are not limitedto, materials that enable the release of the active ingredient to thefood product for a time period ranging from about 12 hours or less, suchas from about 5 seconds to about 12 hours, from about 5 seconds to about10 hours, from about 10 seconds to about 8 hours, from about 15 secondsto about 6 hours, from about 20 seconds to about 4 hours, from about 25seconds to about 2 hours, from about 5 seconds to about 1 hour, fromabout 10 seconds to about 45 minutes, from about 15 seconds to about 30minutes, from about 20 seconds to about 15 minutes, from about 25seconds to about 5 minutes, from about 5 seconds to about 1 minute, fromabout 5 seconds to about 30 seconds, from about 5 seconds to about 15seconds, and from about 5 seconds to about 10 seconds.

The chamber may also comprise one or more holes or apertures. Theapertures may have any shape, and may have a size ranging from about 2mm to about 2 cm, and from about 2.5 mm to about 1.5 cm, from about 5 mmto about 1.5 cm, from about 7.5 mm to about 1.25 cm, from about 10 mm toabout 1 cm, from about 15 mm to about 0.75 cm, from about 20 mm to about0.5 cm, and from about 25 mm to about 0.25 cm. In addition, theapertures may be in any location on the chamber material, but typically,the apertures are located at the base, the lid, the sides, or acombination thereof on the chamber. The apertures allow for introductionof treatment to the chamber and/or release of treatment from thechamber.

Upon introduction of compound treatment into or onto the chamber, theapertures permit uniform distribution of treatment vapor, gas, or fogparticles throughout the chamber. The apertures also allow for fulldrainage, venting, and/or release of the unused portion of the treatmentor treatment carrier from the chamber. Unused treatment and/or treatmentcarrier may be recycled to treat subsequent and/or additionalcontainers, materials, or chambers.

An illustrative example of a product of the method described herein isone or more benzoxaborole-treated PET clamshell, such as a plurality ofPET clamshells. PET clamshells are commonly used to transportstrawberries and other soft fruits. Therefore, a benzoxaborole-treatedPET clamshell would provide the greatest protection to the fruitcontained therein since the active ingredient is coated on the surfacesof the primary packaging of the fruit. Primary protection of the fruitcould also occur via treatment of a material contained within the PETclamshell, such as a liquid-absorbing material in the form of a tag, apad, or other embodiments described herein. Secondary protection of thefruit would occur by applying the active ingredient to a liner, a box, abag, a wrap or other packaging material in which the primary chambersare placed for storage or transport.

Methods of Administering Benzoxaborole Compounds

The present disclosure is directed to methods of uniformly treating foodproducts by providing antimicrobial protection to food, such as plants,crops, and meats. The present methods are directed to large-scaletreatment of food packaging materials to uniformly protect plants fromplant pathogens and microorganisms that cause food decay. Morespecifically, plant pathogens that inhibit, reduce, or compromise foodfreshness may be treated, prevented, or eradicated by the methodsdescribed herein.

Exemplary, microorganisms encompassed by the present disclosure include,but are not limited to, Botrytis cinerea, Mucor piriformis, Fusariumsambucinum, Aspergillus brasiliensis, and Peniciliium expansum.Additional pathogens encompassed by the present invention include, butare not limited to Acremonium spp., Albugo spp., Alternaria spp.,Ascochyta spp., Aspergillus spp., Botryodiplodia spp., Botryospheriaspp., Botrytis spp., Byssochlamys spp., Candida spp., Cephalosporiumspp., Ceratocystis spp., Cercospora spp., Chalara spp., Cladosporiumspp., Colletotrichum spp., Cryptosporiopsis spp., Cylindrocarpon spp.,Debaryomyces spp., Diaporthe spp., Didymella spp., Diplodia spp.,Dothiorella spp., Elsinoe spp., Fusarium spp., Geotrichum spp.,Gloeosporium spp., Glomerella spp., Helminthosporium spp., Khuskia spp.,Lasiodiplodia spp., Macrophoma spp., Macrophomina spp., Microdochiumspp., Monilinia spp., Monilochaethes spp., Mucor spp., Mycocentrosporaspp., Mycosphaerella spp., Nectria spp., Neofabraea spp., Nigrosporaspp., Penicillium spp., Peronophythora spp., Peronospora spp.,Pestalotiopsis spp., Pezicula spp., Phacidiopycnis spp., Phoma spp.,Phomopsis spp., Phyllosticta spp., Phytophthora spp., Polyscytalum spp.,Pseudocercospora spp., Pyricularia spp., Pythium spp., Rhizoctonia spp.,Rhizopus spp., Sclerotium spp., Sclerotinia spp., Septoria spp.,Sphaceloma spp., Sphaeropsis spp., Stemphyllium spp., Stilbella spp.,Thielaviopsis spp., Thyronectria spp., Trachysphaera spp., Uromycesspp., Ustilago spp., Venturia spp., and Verticillium spp., and bacterialpathogens, such as Bacillus spp., Campylobacter spp., Clavibacter spp.,Clostridium spp., Erwinia spp., Escherichia spp., Lactobacillus spp.,Leuconostoc spp., Listeria spp., Pantoea spp., Pectobacterium spp.,Pseudomonas spp., Ralstonia spp., Salmonella spp., Shigella spp.,Staphylococcus spp., Vibrio spp., Xanthomonas spp., and Yersinia spp.

Benzoxaborole treatments may be applied, administered, or coated on theinside or the outside of a chamber or packaging material. When thebenzoxaborole treatment is in any form, but particularly in liquid,spray, vapor, or gas form, a drying step may be provided in the presentmethod that allows excess treatment carrier to dry. This step may alsoproduce a residue of the active benzoxaborole ingredient at the properlevels of efficacy on the surface of the food packaging material inorder to provide extended antimicrobial control and inhibition forplants, crops, and meats contained therein.

Any food product, including plants, crops, or meats, may be treatedusing the present method. Minimally-processed packaged products (e.g.,packaged vegetables, fruits, or meats) may also be treated with themethod described herein. Horticultural crops of the present methodinclude, but are not limited to, vegetable crops, fruit crops, ediblenuts, flowers and ornamental crops, nursery crops, aromatic crops, andmedicinal crops.

Plants and agricultural crops in any production cycle may be used in themethod of the present application. For example, post-harvest plants andcrops may be treated during field packing, palletization, in-box,storage, and throughout the distribution network. Further, plants beingtransported by any mode, including, but not limited to local vehicles,transport trailers, marine containers, aircraft containers, etc. may betreated using the method described herein.

Ideally, the chamber or plurality of chambers of the present disclosureare treated prior to use during food field-packaging of plants, meats,or crops, such as soft fruits. For example, treated chambers may belocated, stored, and/or kept at the site of clamshell manufacturers, atthe central facility of farmers or ranchers, or in a portable unit forimmediate in-field treatments. Additionally, treated chambers ormaterials may be provided to a food producer directly from themanufacturer, wherein the manufacturer has previously appliedbenzoxaborole to the surface of the packaging. Alternatively, a foodproducer may independently use a method, machine, or instrument to treatcontainers or packaging materials with the benzoxaborole compositions asdescribed herein.

Large-scale treatment of food product packaging materials and chambersare comprised in the methods of the present disclosure. Large-scaletreatment comprises treatment of chambers or a plurality of chambers inmass, and typically for commercial and/or industrial use. For example,the methods of the present disclosure may comprise treating a pluralityof chambers with the benzoxaborole active ingredient described herein.Large-scale treatment methods of the present disclosure may occur beforethe chamber has been formed (i.e., preformation), during formation ofthe chamber (i.e., formation), and after the chamber has been formed(i.e., postformation). Formation is the process of forming or producingone or more chambers of the present disclosure, which may includethermoforming.

Preformation treatment of the chamber comprises contacting a packagingmaterial, such as plastic, that will be formed into the chamber with anactive ingredient of the present disclosure (e.g., benzoxaborole) priorto the beginning of the formation process. Formation comprises treatmentof the packaging material of the chamber with the active ingredientafter the formation process has started. Postformation treatment of thechamber occurs when the packaging material has been formed into achamber and the chamber is then treated with the active ingredient. Forexample, the chambers may be treated with the active ingredient priorto, during, or after formation of the chamber using methods including,but not limited to, dipping, drenching, spraying, painting, vaporizing,and/or sublimation.

One embodiment of the large-scale method described herein comprises useof a printer to print the active compound treatment on the lining ormaterial of a plurality of chambers. Another embodiment of thelarge-scale treatment method comprises spraying the plurality ofchambers with the active compound treatment. A further embodiment of thelarge-scale treatment method comprises dipping a plurality of chambersinto a vat of active compound treatment and removing the treatedchambers to dry onto the chamber material. Additionally, fogging orspraying a plurality of chambers with the active compound duringmanufacturing in an industrial-sized device is also an embodiment of thelarge-scale method of the present disclosure.

Food products may or may not be inside of the chamber during applicationof the benzoxaborole treatment. If the food product is already insidethe chamber, treatment of the chamber with the active ingredient may beapplied while the chamber is open, closed, or sealed. Typically,however, after the treated chambers are produced, food products, such asplants, crops, or meats, may be manually or robotically (e.g., by amachine) placed in the treated chamber in preparation for antimicrobialtreatment of the food.

The proximity and/or distance between the emitting source of activeingredient and the food product is critical. Notably, the distance fromthe benzoxaborole coated surfaces of the packaging material or chamberand the food product is inversely related to the efficacy ofantimicrobial protection of the food. In other words, the greater thedistance between the coated/treated surfaces of the material or chamberand the food product, the lesser is the level of antimicrobialprotection conveyed to the treated food product, including plants,crops, or meat.

Related to this property, treated surfaces of a chamber or packagingmaterial, such as a bag, a box, a wrap, a liner, or other packagingmaterial that is placed over an entire pallet of clamshells of foodproducts may be less effective in delivering the active ingredient tothe food than a treated surface that is immediately adjacent to or incontact with the plant, crop, or meat food products, such as theclamshell surfaces themselves. Therefore, having the treatment coateddirectly on a surface or imbedded into the individual product chamberthat is the primary packaging material, meaning the first layer ofpackaging of the food product, provides the greatest antimicrobialprotection to the food product. Similarly, treating the internal surfaceof the primary packaging material, such as a chamber, provides evengreater antimicrobial effect than treating an external surface of thechamber. Therefore, the distance between the emitting source of theactive ingredient and the food product should be minimized for bestresults.

More specifically, the distance between the emitting source and the foodproduct should remain less than about 6 feet. In an illustrativeembodiment, the distance between the emitting source and the foodproduct ranges from about 0.1 inches to about 6 feet, from about 0.5inches to about 5 feet, from about 1 inch to about 4 feet, from about1.5 inches to about 3 feet, from about 2 inches to about 2 feet, fromabout 0.5 inches to about 12 inches, from about 1 inch to about 24inches, from about 0.5 inches to about 6 inches, from about 0.5 inchesto about 5 inches, from about 0.5 inches to about 4 inches, from about0.5 inches to about 3 inches, from about 0.5 inches to about 2 inches,and from about 0.5 inches to about 1 inch. Close proximity of theemitting source of the benzoxaborole active ingredient applied to theprimary PET clamshell ensures that the food product is exposed to theactive ingredient which inhibits microorganisms that may infect thefood.

An exemplary embodiment of the method described herein comprisesvaporizing or subliming the benzoxaborole compound or molecule into agaseous form. The benzoxaborole compound gas or vapor may be at anyconcentration that allows the compound or molecule to adhere to thechamber (e.g., a clamshell) or packaging surfaces prior to taking thechamber or material into a field for the harvest operation.

For example, the benzoxaborole compound vapor or gas may be effectivelyadministered to a chamber at a concentration ranging from about 0.1mg/chamber to about 10 mg/chamber, from about 0.1 mg/chamber to about 8mg/chamber, from about 0.1 mg/chamber to about 7 mg/chamber, from about0.1 mg/chamber to about 6 mg/chamber, from about 0.1 mg/chamber to about5.5 mg/chamber, from about 0.1 mg/chamber to about 5 mg/chamber, fromabout 0.1 mg/chamber to about 4 mg/chamber, from about 0.1 mg/chamber toabout 3.5 mg/chamber, from about 0.1 mg/chamber to about 3.2 mg/chamber,from about 0.1 mg/chamber to about 2 mg/chamber, from about 0.1mg/chamber to about 1.5 mg/chamber, from about 0.1 mg/chamber to about 1mg/chamber, from about 0.1 mg/chamber to about 0.35 mg/chamber, fromabout 0.1 mg/chamber to about 0.32 mg/chamber, from about 0.1 mg/chamberto about 0.25 mg/chamber, from about 0.1 mg/chamber to about 0.22mg/chamber, 0.2 mg/chamber to about 8 mg/chamber, from about 0.2mg/chamber to about 7 mg/chamber, from about 0.2 mg/chamber to about 6mg/chamber, from about 0.2 mg/chamber to about 5.5 mg/chamber, fromabout 0.2 mg/chamber to about 5 mg/chamber, from about 0.2 mg/chamber toabout 4 mg/chamber, from about 0.2 mg/chamber to about 3.5 mg/chamber,from about 0.2 mg/chamber to about 3.2 mg/chamber, from about 0.2mg/chamber to about 2 mg/chamber, from about 0.2 mg/chamber to about 1.5mg/chamber, from about 0.2 mg/chamber to about 1 mg/chamber, from about0.2 mg/chamber to about 0.35 mg/chamber, from about 0.2 mg/chamber toabout 0.32 mg/chamber, from about 0.2 mg/chamber to about 0.25mg/chamber, from about 0.2 mg/chamber to about 0.22 mg/chamber, 0.2mg/chamber to about 8 mg/chamber, from about 0.3 mg/chamber to about 7mg/chamber, from about 0.3 mg/chamber to about 6 mg/chamber, from about0.3 mg/chamber to about 5.5 mg/chamber, from about 0.3 mg/chamber toabout 5 mg/chamber, from about 0.3 mg/chamber to about 4 mg/chamber,from about 0.3 mg/chamber to about 3.5 mg/chamber, from about 0.3mg/chamber to about 3.2 mg/chamber, from about 0.3 mg/chamber to about 2mg/chamber, from about 0.3 mg/chamber to about 1.5 mg/chamber, fromabout 0.3 mg/chamber to about 1 mg/chamber, from about 0.3 mg/chamber toabout 0.35 mg/chamber, from about 0.3 mg/chamber to about 0.32mg/chamber, and at about 0.2 mg/chamber, 0.316 mg/chamber, 1 mg/chamber,3.16 mg/chamber, and 5 mg/chamber.

Alternatively, the benzoxaborole compound may be prepared as a liquidformulation. Preparing a liquid composition of the benzoxaboroletreatment requires mixing the benzoxaborole compound with a liquidcarrier, such as a solvent or water or combination thereof. Once theliquid treatment is prepared, the chamber or material is drenched orflooded with the liquid treatment, such that the liquid treatmenttouches all internal surfaces. Most of the liquid treatment carrier maythen be drained from an aperture or hole in the chamber or material.After the carrier is substantially removed from the chamber, residue ofthe active ingredient (i.e., benzoxaborole) in the remaining liquid isallowed to dry, such as at room temperature (e.g., about 21° C. to about23° C.).

Drying of the treatment composition, including the liquid carrier, mayoccur instantaneously or within seconds (secs). In particular, thehigher the concentration of active ingredient (i.e., benzoxaborole) andthe lower the volume of compound treatment, the less time is requiredfor drying the treatment composition onto the chamber. For example,drying time of the treatment composition onto the chamber material mayrange from about 0.1 secs to about 60 secs, from about 0.2 secs to about45 secs, from about 0.3 secs to about 30 secs, from about 0.4 secs toabout 20 sec, from about 0.5 secs to about 15 secs, from about 1 sec toabout 10 secs, from about 5 secs to about 60 secs, and at about 5 secs.Once dried, the active benzoxaborole ingredient is coated onto thesurface of the chamber to provide immediate antimicrobial protection tofood, such as plants, crops, or meats placed therein.

As previously mentioned, a liquid benzoxaborole treatment may beadministered to a fruit or a vegetable clamshell, such that all internalsurfaces of the chamber come into contact with the active ingredient.After draining the treatment carrier (e.g., water or solvent) from theclamshell, warm air (i.e., room temperature) is applied to dry theremaining liquid on the clamshells. The treated clamshells comprisingthe coated surfaces and active ingredient may then go immediately to thefield for picking operations. Food, such as fresh fruits and berriesplaced within the treated clamshells, is protected from diseasemicroorganisms by the volatile active ingredient (i.e., benzoxaborole)emitted from the surface of the clamshells from the time the fruit isplaced inside of the chamber.

In yet another embodiment of the present method, the benzoxaboroleactive ingredient may be administered by fogging as a fine mist into asuitable chamber. The compound may be fogged using any cold, thermal,ultrasonic, or similar fogging-based technologies. The micron particlesof water, solvent, or other carriers in the formulations may assistdistribution and deposition of the benzoxaborole particles on thechamber surfaces. Upon drying, the method will result in a thin coatingof active ingredient on the chamber surfaces. This thin coating ofbenzoxaborole will volatilize over time, and uniformly protect the foodcontents of the chamber from pathogenic infection and decay.

Accordingly, the method described herein provides a large-scaleadministration of an antimicrobial agent, such as benzoxaborole, to thepackaging surfaces of food product chambers. Importantly, the presentlydescribed method enables greater uniformity and consistency ofapplication of the active benzoxaborole treatment composition to thefood packaging materials. Ultimately, the present large-scale method ofuniformly treating food chambers results in a significantly extendedtime period (i.e., up to about 31 days or a full month) of antimicrobialprotection of the food product comprised within the treated chambers.

EXAMPLES

Illustrative embodiments of the methods of the present disclosure areprovided herein by way of examples. While the concepts and technology ofthe present disclosure are susceptible to broad application, variousmodifications, and alternative forms, specific embodiments will bedescribed here in detail. It should be understood, however, that thereis no intent to limit the concepts of the present disclosure to theparticular forms disclosed, but on the contrary, the intention is tocover all modifications, equivalents, and alternatives consistent withthe present disclosure and the appended claims.

The following experiments were used to determine the effect of differentconcentrations of benzoxaborole compounds when administered onto thesurfaces of packaging materials or chambers by various applicationtechniques. In the following experiments, benzoxaborole treatmentcompositions are applied to chambers such that food products may beexposed to the antimicrobial treatment for an initial time period. Forexample, food products may be exposed to the benzoxaborole treatment onthe surfaces of the treated chamber for the initial time period rangingfrom less than 1 day to about 8 days, and at about 5 days. Treatmenttemperatures of the chamber during the initial time period ranged fromabout 0.5° C. to about 5° C., and at about 1° C.

After the initial time period in which the food is exposed to the activeingredient of the treated chamber, the chamber may be unsealed (ifpreviously sealed), and allowed to vent for a secondary time period. Thefood may remain in the chamber for the secondary time period rangingfrom about 1 day to about 8 days, and at about 6 days. The temperatureof the chamber during the second time period remains at roomtemperature, which ranges from about 20° C. to about 23° C., and atabout 21° C.

After expiration of the secondary time period, inhibition of plantpathogens and infection may be assessed. For example, in vitro samplesmay have the growth of the microorganism or pathogen on agar or in mediaassessed, evaluated, and compared to a control sample where nobenzoxaborole treatment was administered. Similarly, in vivo samples mayhave the severity and incidence of pathogenic disease assessed,evaluated, and compared to a control sample where no benzoxaboroletreatment was administered or different treatment conditions wereapplied.

Example 1: Benzoxaborole Compound Treatment of Fruit Clamshells by

Spraying, Painting, and Sublimation (In Vivo)

An in vivo assay was used to evaluate the ability of Compound A tovolatilize from a clamshell chamber and control pathogenic infectionwhen applied by various techniques. Multiple empty 1-lb PET clamshells(ProducePackaging.com, #036QT) were placed inside triplicate air-tight36 L chamber (Fisher Scientific, Catalogue #08-642-23C) fitted with abulkhead septum port (Swagelok, SS-401-61, Solon, Ohio).

An appropriate amount of Compound A, to achieve a final treatment rateof 5 mg per clamshell (i.e., 5 mg/clamshell), was dissolved in acetoneand 100 μL of the solution was pipetted into a small glass tube. Thetube was then placed inside a pre-heated sublimation device (0.5″ OD by6″ long thermostatically healed copper tube mounted to a 2 L/minaquarium pump) set at 60° C. for 1 minute to allow the acetone toevaporate. Compound A was then introduced into the cabinets of thechamber through the bulkhead port containing the clamshells by using thesublimation device set at 180° C. Compound A headspace was permitted toequilibrate overnight at 21° C.

Five milligrams of Compound A was dissolved in 1 ml of ethanol prior tobeing uniformly administered to the interior of the clamshell bypainting or spraying, and then dried for 5 minutes. After coating theclamshells with Compound A using various application techniques (i.e.,sublimation, spraying, or painting), eight ethanol-washed strawberrieswere placed in the clamshell with stem end facing downwards. Eachstrawberry fruit was wounded using a T15 screwdriver tip to a uniformdepth of eight mm (8 mm). Each fruit wound was inoculated with 20 μL of1×10⁵ spores/ml pathogen spore suspension of Botrytis cinerea, which isa fungal pathogen known to cause gray mold infection of fruits, such asgrapes and strawberries. Uninoculated strawberries were removed fromtheir commercial package, and directly transferred into treatedclamshells without any washing or inoculation steps.

Treated clamshells lids were closed, and then placed at 1° C. for aninitial time period of 5 days. Clamshells were then removed from lowtemperature, and held for a second time period of 6 days at roomtemperature where the point of inoculation on the strawberry fruits wasassessed for indication of disease incidence reported as a percentage(%). Severity of disease incidence was also reported. Disease severitywas rated on a scale ranging from 0 to 4, where “0” indicated no diseaseseverity, “1” indicated minimal disease severity, “2” indicated mediumdisease severity, “3” indicated high disease severity, and “4” indicatedexceptionally high disease severity.

The outcome of this in vivo experiment is summarized in Table 1. Resultsdemonstrate good in vivo antimicrobial activity of Compound A against B.cinerea, with a reduction in disease incidence and severity with allthree application techniques (i.e., painting, spraying, andsublimation). In particular, each method of treating clamshells showedsignificant inhibition of gray mold incidence and severity instrawberries as compared to control. More specifically, on Days 1-6, thepercentage of gray mold incidence increased from 30.5% to 100% and 0% toa maximum of 0.9% in control fruits and treated inoculated fruits,respectively. Even in uninoculated fruit, the percentage of gray moldincidence increased from 1.5% to 100% and 0% to a maximum of 21.7% incontrol fruits and treated fruits, respectively. In both inoculated anduninoculated fruits, the spraying technique was comparable to or betterthan painting or sublimation in minimizing the incidence or severity ofgray mold. Ultimately, treating the clamshells with benzoxaboroleCompound A significantly inhibited the growth of B. cinerea in thestrawberries and preserved the freshness of the fruit for at least 3days longer than the untreated strawberries.

Example 2: Benzoxaborole Compound Treatment of Clamshells ContainingAgar Plates by Spraying, Painting, and Sublimation (In Vitro)

An in vitro assay was used to evaluate the ability of Compound A tovolatilize from a clamshell to control fungal pathogenic infection whenapplied to the clamshell by various application techniques. Multipleempty 1-lb PET clamshells (ProducePackaging.com, #036QT) were placedinside triplicate air-tight 36 L chamber (Fisher Scientific, Catalogue#08-642-23C) fitted with a bulkhead septum port (Swagelok, SS-401-61,Solon, Ohio).

An appropriate amount of Compound A to achieve a final treatment rate of5 mg/clamshell, 1 mg/clamshell, or 0.2 mg/clamshell, was dissolved inacetone and 100 μL of the solution was pipetted into a small glass tube.The tube was then placed inside a pre-heated sublimation device (0.5″ ODby 6″ long thermostatically healed copper tube mounted to a 2 L/minaquarium pump) set at 60° C. for 1 minute to allow the acetone toevaporate. Compound A was then introduced into the cabinets of thechamber through the bulkhead port containing the clamshells by using thesublimation device set at 180° C. Compound A headspace was permitted toequilibrate overnight at 21° C.

For paint and spray applications, 5 mg of Compound A was dissolved in 1ml of ethanol prior to uniformly painting or spraying the interior ofthe clamshell. After spraying or painting, the clamshell was thenpermitted to dry for 5 minutes. After coating the clamshells withCompound A using various applications (i.e., sublimation, spraying, orpainting), 10-cm Petri plates containing half strength Potato DextroseAgar were inoculated with 1 μL of 1×10⁵ spores/ml Botrytis cinerea sporesuspension. The inoculated petri plates were then sealed with abreathable film (AeraSeal; P/N: B-100, Excel Scientific, Victorville,Calif.), and placed inside the treated clamshell.

To determine the period of time coated packaging could release effectivelevels of the active ingredient, treated clamshells containing theinoculated plates were then placed inside a 2.55 L SnapWare airtightcontainer (Model #109842) for three (3) days at 21° C. (Series I). Afterincubation, plates were removed and cultures were evaluated for percentgrowth relative to a control based on measurement of fungal colonydiameter (mm).

TABLE 1 Comparison of in-clamshell Compound A application techniques tocontrol growth of Botrytis cinerea inoculated strawberries as comparedto uninoculated fruit. Gray Mold Incidence (%) Inoculated Fruit Day 0Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Control 30.5 62.1 84.3 93.5 100.0100.0 100.0 Paint 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Spray 0.0 0.0 0.0 0.0 0.00.0 0.0 Sublimation 0.0 0.0 0.0 0.0 0.0 0.9 0.9 Gray Mold Severity (0-4)Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Control 0.2 0.4 1.1 2.1 2.64.0 4.0 Paint 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Spray 0.0 0.0 0.0 0.0 0.0 0.00.0 Sublimation 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Gray Mold Incidence (%)Uninoculated Fruit Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Control 1.57.1 39.9 96.7 96.7 100.0 100.0 Paint 0.0 0.3 3.9 5.1 7.4 17.0 21.7 Spray0.0 0.6 2.1 4.8 6.0 17.9 19.9 Sublimation 0.0 0.3 1.5 3.3 5.7 16.7 20.2Gray Mold Severity (0-4) Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6Control 0.3 0.9 1.5 2.8 3.5 4.0 4.0 Paint 0.0 0.1 0.5 0.8 0.9 1.1 1.1Spray 0.0 0.2 0.3 0.5 0.7 1.2 1.3 Sublimation 0.0 0.1 0.3 0.7 0.9 1.11.3

Meanwhile, freshly prepared inoculated plates were placed inside theclamshell for an additional three (3) days of incubation (Series 2).This process, where freshly prepared inoculated plates were placedinside the clamshell, was repeated twice more, to produce a 12 full dayevaluation of 4 series of 3 days each. The outcome of this in vitroexperiment is summarized in Table 2. Results demonstrate good volatilein vitro antimicrobial activity of Compound A against Botrytis cinereawith all three application techniques (i.e., painting, spraying, andsublimation). In addition, greater inhibition of pathogenic growth wasobserved at higher treatment rates.

In particular, each technique of treating clamshells showed significantinhibition of mycelial growth. More specifically, administration of 5mg/clamshell of benzoxaborole treatment by all three techniques wereeffective to completely inhibit mycelial cell growth the first six days(Table 2). In the remaining six days, sublimation most effectivelyinhibited mycelial growth (84.4%), followed by spraying (64.9%), andpainting (44.7%). At lower treatment concentrations (i.e., 1mg/clamshell and 0.2 mg/clamshell), all of the treatment techniquesfailed to inhibit mycelial cell growth by Days 6 to 9. Ultimately,treating the clamshells with benzoxaborole Compound A significantlyinhibited the growth of B. cinerea inoculated on agar plates placedtherein for time periods typical of fruit storage by the supply chainand consumers.

TABLE 2 Comparison of treatment techniques of Compound A to volatilizefrom clamshells and provide in vitro inhibition of Botrytis cinereaMycelial Growth Inhibition (%) Rate Series 1 Series 2 Series 3 Series 4(mg/ (0 to (3 to (6 to (9 to Method clamshell) 3 days) 6 days) 9 days)12 days) Sublimation 5 100.0 100.0 83.1 84.4 1 90.1 0.0 0.0 . . . 0.290.9 0.0 0.0 . . . Paint 5 100.0 100.0 57.5 44.7 1 100.0 18.8 0.0 — 0.23.4 0.0 0.0 — Spray 5 100.0 100.0 61.2 64.9 1 100.0 18.6 0.0 . . . 0.281.2 0.0 0.0

Example 3: Dose Response of Benzoxaborole Compound Treatment of FruitClamshells by Vapor-Coating (In Vivo)

This in vivo assay was used to evaluate the ability of vaporizedCompound A to volatilize from a clamshell in order to control or inhibitpathogenic microorganisms. This experiment was conducted exactly asdescribed in Example 1, with a few exceptions. After equilibrating theclamshell overnight at 21° C., Compound A was administered to theclamshell as a vapor only. In addition, Compound A was vapor-coated ontothe surface of the clamshell at a final treatment rate of 3.16mg/clamshell, 1 mg/clamshell, or 0.316 mg/clamshell. After vapor-coatingapplication, Compound A headspace was permitted to equilibrate overnightat 21° C. Strawberries were inoculated with Botrytis cinerea, placedwithin the clamshells for an initial time period, and assessed fordisease incidence and severity over a second time period as described inExample 1.

The outcome of this in vivo experiment is summarized in Table 3. Resultsdemonstrate good in vivo antimicrobial activity of Compound A against B.cinerea, with a greater reduction in disease incidence and severityobserved at higher treatment rates.

In particular, each concentration of active ingredient on treatedclamshells showed inhibition of gray mold severity in inoculatedstrawberries as compared to control (see Table 3). More specifically, onDays 1-6, the percentage of gray mold severity increased from 0.8 to 4.0in control fruits as compared to 0 to 0.2 and 0 to 0.7 for inoculatedfruits treated with 3.16 mg/clamshell or 1 mg/clamshell of Compound A,respectively. At the lower treatment concentration of 0.316mg/clamshell, the percentage of gray mold severity increased from 0.4 to3.9. Ultimately, treating the clamshells with different concentrationsof benzoxaborole Compound A significantly inhibited the growth of B.cinerea in the strawberries in a dose dependent manner.

TABLE 3 Dose response of in-clamshell Compound A applied by sublimationto control growth of Botrytis cinerea inoculated on strawberries.Inoculated Fruit Rate Gray Mold Severity (0-4) (mg per clamshell) Day 1Day 2 Day 3 Day 4 Day 5 Day 6 3.16 0.0 0.0 0.0 0.0 0.1 0.2 1.00 0.0 0.00.0 0.1 0.4 0.7 0.316 0.4 1.3 2.2 3.4 3.8 3.9 Control 0.8 1.8 2.7 3.64.0 4.0

Example 4: Dose Response of Benzoxaborole Compound Treatment of Fruit

Clamshells by Spraying (In Vivo)

This in vivo assay was used to evaluate the ability of differentconcentrations of Compound A to volatilize from clamshell in order tocontrol or inhibit fruit infection by pathogenic microorganism, Botrytiscinerea. This experiment was conducted exactly as described in Example1, with a few exceptions. After equilibrating the clamshell overnight at21° C., Compound A was administered to the clamshells by spraying only.Compound A was sprayed onto the surface of the clamshells at a finaltreatment rate of 5 mg/clamshell or 1 mg/clamshell. The clamshells werethen permitted to dry for 5 minutes. Strawberries were inoculated withBotrytis cinerea, placed within the clamshells for the initial timeperiod (i.e., 5 days), and assessed for disease incidence and severityover the second time period (i.e., 6 days) as described in Example 1.

The outcome of this in vivo experiment is summarized in Table 4. Resultsdemonstrate good in vivo antimicrobial activity of Compound A against B.cinerea, with a reduction in disease incidence and severity with both 5mg/clamshell and 1 mg/clamshell concentrations applied by spraying. Inparticular, each concentration of treating clamshells showed inhibitionof gray mold incidence and severity in strawberries as compared tocontrol. On Days 1-6, the percentage of gray mold incidence increasedfrom 6.3% to 100% in control inoculated fruits, while there was nogrowth in 5 mg/clamshell-treated inoculated fruits. Even the 1mg/clamshell-treated inoculated fruits inhibited gray mold incidence toa 52.5% maximum.

In uninoculated fruit, the percentage of gray mold incidence increasedfrom 0.5% to 78.5% in control fruits, and similarly, from 0% to 100% in1 mg/clamshell-treated uninoculated fruits. However, the percentage ofgray mold incidence only increased from 0.5% to 78.5% in 5mg/clamshell-treated uninoculated fruits.

In addition, each concentration of active ingredient on treatedclamshells showed significant inhibition of gray mold severity ininoculated strawberries as compared to control (see Table 4). Morespecifically, on Days 1-6, the degree of gray mold severity increasedfrom 0.0 to 4.0 in control fruits as compared to 0 to 1.5 in 1mg/clamshell-treated inoculated fruits and no growth in 5mg/clamshell-treated inoculated fruits. For uninoculated fruits, bothcontrol and 1 mg/clamshell-treated fruits showed a gray mold severitylevel of 4.0 by Day 4, while the 5 mg/clamshell-treated uninoculatedfruits only showed a gray mold severity level of 3.5 on Day 6.Ultimately, these data demonstrate that treating clamshells withdifferent concentrations of benzoxaborole Compound A significantlyinhibited the gray mold infection of B. cinerea inoculated instrawberries in a dose dependent manner.

TABLE 4 Dose Response of in-clamshell Compound A application techniquesto control growth of Botrytis cinerea inoculated in strawberries ascompared to uninoculated strawberries. Inoculated Fruit Gray MoldIncidence (%) Rate (mg/clamshell) Day 0 Day 1 Day 2 Day 3 Day 4 Day 5Day 6 5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 1 0.0 0.0 1.3 3.8 11.3 36.3 52.5Control 6.3 72.5 100.0 100.0 100.0 100.0 100.0 Gray Mold Severity (0-4)Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 5 0.0 0.0 0.0 0.0 0.0 0.0 0.01 0.0 0.0 0.0 0.1 0.2 0.6 1.5 Control 0.0 0.7 1.4 2.7 3.6 3.8 4.0Uninoculated Fruit Gray Mold Incidence (%) Rate (mg/clamshell) Day 0 Day1 Day 2 Day 3 Day 4 Day 5 Day 6 5 0.5 0.5 0.9 3.3 16.2 31.9 78.5 1 0.01.9 21.6 89.5 100.0 100.0 100.0 Control 1.0 6.0 46.0 100.0 100.0 100.0100.0 Gray Mold Severity (0-4) Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 65 0.1 0.1 0.3 0.5 1.0 1.4 3.5 1 0.0 0.5 1.8 3.6 4.0 4.0 4.0 Control 0.31.0 2.9 4.0 4.0 4.0 4.0

Example 5: Benzoxaborole Compound Treatment of Fruit Clamshells by

Sublimation (In Vivo)

This in vivo assay was used to evaluate the ability of Compound A tovolatilize from a clamshell in order to control or inhibit fruitinfection by pathogenic microorganism, Botrytis cinerea. This experimentwas conducted exactly as described in Example 1, with a few exceptions.After equilibrating the clamshell overnight at 21° C., Compound A wasadministered to the clamshell by sublimation only. Compound A wassublimed onto the surface of the clamshell at a final treatment rate of5 mg/clamshell. Strawberries were inoculated with Botrytis cinerea,placed within the clamshell for an initial time period of 6 days, andassessed for disease incidence and severity over a second time period of7 days as described in Example 1.

The outcome of this in vivo experiment is summarized in Table 5. Resultsdemonstrate good in vivo antimicrobial activity of Compound A against B.cinerea, with a reduction in disease incidence and severity with the 5mg/clamshell concentrations applied by sublimation. In particular, bothinoculated and uninoculated 5 mg/clamshell-treated clamshells showedinhibition of gray mold incidence and severity in strawberries ascompared to control. On Days 1-7, the percentage of gray mold incidenceincreased from 14.6% to 100% in control inoculated fruits, while therewas a maximum of 18.8% of gray mold incidence in 5 mg/clamshell-treatedinoculated fruits observed on Day 6.

In uninoculated fruit, the percentage of gray mold incidence increasedfrom 0% to 100% in control fruits, however, the percentage of gray moldincidence only increased to 54.1% in 5 mg/clamshell-treated uninoculatedfruits.

In addition, the 5 mg/clamshell concentration of active ingredient onsublimation-treated clamshells showed significant inhibition of graymold severity in inoculated strawberries as compared to control (seeTable 5). More specifically, on Days 1-7, the degree of gray moldseverity increased from 0.1 to 4.0 in control inoculated fruits ascompared to 0 to 0.3 in 5 mg/clamshell-treated inoculated fruits. Foruninoculated fruits, the control fruits showed a gray mold severitylevel of 4.0 by Day 5, while the 5 mg/clamshell-treated uninoculatedfruits only showed a gray mold severity level level of 1.6 on Day 7.Ultimately, this data demonstrate that treating clamshells with 5mg/clamshell of benzoxaborole Compound A inhibited the gray moldinfection of B. cinerea inoculated in strawberries.

TABLE 5 Ability of Compound A applied to clamshells by sublimation tocontrol growth of Botrytis cinerea inoculated in strawberries.Inoculated Fruit Gray Mold Incidence (%) Treatment Day 0 Day 1 Day 2 Day3 Day 4 Day 5 Day 6 Day 7 5 mg Compound A 0.0 0.0 0.0 0.0 0.0 16.7 18.816.7 Control 14.6 52.1 91.7 95.8 97.9 100.0 100.0 100.0 Gray MoldSeverity (0-4) Treatment Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 75 mg Compound A 0.0 0.0 0.0 0.0 0.0 0.1 0.2 0.3 Control 0.1 0.4 0.9 2.03.0 4.0 4.0 4.0 Uninoculated Fruit Gray Mold Incidence (%) Treatment Day0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 5 mg Compound A 0.0 2.6 0.126.3 34.1 40.0 43.3 54.1 Control 0.0 23.0 0.6 95.2 100.0 100.0 100.0100.0 Gray Mold Severity (0-4) Treatment Day 0 Day 1 Day 2 Day 3 Day 4Day 5 Day 6 Day 7 5 mg Compound A 0.0 0.4 1.0 1.0 1.0 1.3 1.3 1.6Control 0.1 1.0 1.0 2.0 2.8 4.0 4.0 4.0

Example 6: Benzoxaborole Compound Treatment to Different Locations ofFruit

Clamshells by Painting (In Vivo)

This in vivo assay was used to evaluate the ability of Compound A tovolatilize from different locations of a clamshell (i.e., the baseand/or the lid of the clamshell) in order to control or inhibit fruitinfection by pathogenic microorganism, Botrytis cinerea. This experimentwas conducted exactly as described in Example 1, with a few exceptions.Compound A was administered to the clamshell by painting only. 5 mg ofCompound A was painted onto the surface of the base of the clamshell orthe lid of the clamshell (i.e., 5 mg/clamshell treatment rate). 2.5 mgof Compound A was painted onto the base and the lid of the clamshell(for a total of 5 mg/clamshell treatment rate). The clamshell was thenpermitted to dry for 5 minutes. Strawberries were inoculated withBotrytis cinerea, placed within the clamshell for an initial time periodof 5 days, and assessed for disease incidence and severity over a secondtime period of 7 days as described in Example 1.

The outcome of this in vivo experiment is summarized in Table 6. Resultsdemonstrate good in vivo antimicrobial activity of Compound A against B.cinerea, with a reduction in disease incidence and severity with the 5mg/clamshell concentrations applied by painting. In particular, bothinoculated and uninoculated 5 mg/clamshell-treated clamshells showedinhibition of gray mold incidence and severity in strawberries ascompared to control. On Days 1-7, the percentage of gray mold incidenceincreased from 0% to 100% in control inoculated fruits, while there wasa maximum of 18.8% of gray mold incidence in 5 mg/clamshell base-treatedinoculated fruits observed on Day 7. However, inoculated fruits inclamshells painted with 5 mg of Compound A on the lids only or the baseand lids showed no incidence of gray mold even by Day 7.

In uninoculated fruit, the percentage of gray mold incidence increasedfrom 0% to 100% in control fruits, however, the percentage of gray moldincidence only increased to 41.1%, 64.4%, and 52.2% in uninoculatedfruits painted with 5 mg/clamshell on the base only, the lid only, andthe base and lid, respectively.

In addition, the 5 mg/clamshell concentration of active ingredient onpainted clamshells showed significant inhibition of gray mold severityin inoculated strawberries as compared to control (see Table 6). Morespecifically, on Days 1-7, the degree of gray mold severity increasedfrom 0 to 3.6 in control inoculated fruits as compared to 0 to 0.3 in 5mg/clamshell base-treated inoculated fruits. However, inoculated fruitsin clamshells painted with 5 mg of Compound A on the lids only or thebase and lids showed no increase in the severity of gray mold even byDay 7.

For uninoculated fruits, the control fruits showed a gray mold severitylevel of 4.0 by Day 5, while the gray mold severity level level was 1.5,2.5, and 1.5 on Day 7 in uninoculated fruits painted with 5 mg/clamshellon the base only, the lid only, and the base and lid, respectively.Ultimately, these data demonstrate that treating clamshellswith/clamshell of benzoxaborole Compound A significantly inhibited thegray mold infection of B. cinerea irrespective of the location of thetreatment application.

TABLE 6 Ability of Compound A painted on the base, lid, or base and lidof clamshells to control growth of Botrytis cinerea inoculated onstrawberries. Inoculated Fruit Gray Mold Incidence (%) TreatmentLocation Rate (mg) Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Base5 0.0 0.0 12.5 12.5 0.0 0.0 18.8 18.8 Lid 5 0.0 0.0 0.0 0.0 0.0 0.0 0.00.0 Base and Lid 2.5/2.5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Control 0 0.025.0 56.3 68.8 93.8 93.8 93.8 100.0 Gray Mold Severity (0-4) TreatmentLocation Rate (mg) Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Base5 0.0 0.0 0.1 0.1 0.0 0.0 0.2 0.3 Lid 5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0Base and Lid 2.5/2.5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Control 0 0.0 0.10.5 0.8 1.5 2.7 3.3 3.6 Uninoculated Fruit Gray Mold Incidence (%)Treatment Location Rate (mg) Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6Day 7 Base 5 0.0 0.0 0.0 0.0 2.2 10.0 26.7 41.1 Lid 5 0.0 0.0 0.0 1.118.9 27.8 45.6 64.4 Base and Lid 2.5/2.5 0.0 0.0 0.0 1.1 7.8 23.3 33.352.2 Control 0 0.0 1.1 1.1 18.9 88.9 97.8 100.0 100.0 Gray Mold Severity(0-4) Treatment Location Rate (mg) Day 0 Day 1 Day 2 Day 3 Day 4 Day 5Day 6 Day 7 Base 5 0.0 0.0 0.0 0.0 0.3 0.8 1.2 1.5 Lid 5 0.0 0.0 0.0 0.21.2 2.0 2.0 2.5 Base and Lid 2.5/2.5 0.0 0.0 0.0 0.2 0.8 1.5 1.5 1.5Control 0 0.0 0.2 0.2 1.3 3.7 4.0 4.0 4.0

Example 7

For testing activity against fungi pathogens, an in vitro inhibitionassay for volatile antimicrobial compounds is developed using 12-Well (7milliliter (mL) volume per well) microtiter plates. A 3-mL volume offull-strength Potato Dextrose Agar (PDA) is added to each well. Aftercooling, 1 microliter (μL) of 1×10⁶ per mL Botrytis cinerea sporesuspension is spot pipetted to the center of the agar. For the firstexperiment, inoculated plates are allowed to germinate for 5 days at 4°C. For the second experiment, plates are inoculated immediately prior tovolatile fungicide treatment. Small Whatman #1 filter disks (Cat. No.1001-0155) are placed, in duplicate, on the underside of a polyethylenePCR plate sealing film.

TABLE 7 Results of in vitro assay for volatile fungicide Rate ofCompound A Botrytis inhibition % (mg per disk) (in vitro) 1.25 100% 0.63100% 0.31 100% 0.16 100% 0.08 100% 0.04 100% 0.023 100% 0.01 100% 0.005100% 0.0024  85% 0.001  69% 0.0006  46% Control  0%

For determination of the minimum inhibitory concentration (MIC),Compound A (5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole) is dilutedin acetone, and the appropriate amount of compound is added to disks ina dose dependent manner (1.25 to 0.0006 milligrams per disk (mg/disk)).The acetone is permitted to evaporate for 5 minutes. The headspacearound the Botrytis cinerea inoculum is then sealed inside the well bythe film with the adhering disk containing the fungicide. Plates areinverted, placed over the treated disks and sealed to prevent any of thechemical from flaking from the disk and falling onto the inoculatedagar. After 14 days of storage at 4° C., cultures are evaluated forpercent growth relative to control. Regardless of whether the spores hadgerminated for 5 days, or if the treatment commenced soon afterinoculation of the plates (˜15 minutes); there is 100% control of thefungal pathogen down to 0.005 mg.

Experimental results are summarized in Table 1. The results suggest thatCompound A is able to kill Botrytis cinerea spores and inhibit mycelialgrowth at the same concentration. Thus, Compound A shows 100% efficacyin the in vitro inhibition of fungal growth at a rate of 0.005 mg/disk.

Compound B2 (2-(hydroxymethyl)phenylboronic acid cyclic monoester, ades-fluoro analogue of Compound A), is evaluated in a similar manner.The compound is applied to the Whatman filter paper at rates from 0.5 mgto 0.0039 mg/disk. Results show that Compound B2 inhibits 100% Botrytiscinerea at a rate of 0.0078 mg/disk.

Example 8

For testing activity against bacteria pathogens, 12-Well (7 mL volumeper well) microtiter plates are used for the in vitro inhibition assayfor volatile antimicrobial compounds. A 3-mL volume of full-strength LBAgar is added to each well. After cooling, 15 μL of Escherichia coli,adjusted to an optical density of 0.02 to 0.035, and further diluted1/10 is pipetted to the center of the agar and tilted to distributeuniformly. Small Whatman #1 filter disks (Cat. No. 1001-0155) areplaced, in duplicate, on the underside of a polyethylene polymerasechain reaction (PCR) plate sealing film. For determination of theminimum inhibitory concentration (MIC), Compound A is diluted inacetone, and 5 mg of compound is added to the disks. The acetone ispermitted to evaporate for 5 minutes. The headspace around theEscherichia coli inoculum is then sealed inside the well by the filmwith the adhering disk containing the fungicide. Plates are inverted,placed over the treated disks and sealed to prevent any of the chemicalfrom flaking from the disk and falling onto the inoculated agar. After 3days of storage at 4° C., cultures are transferred to 23° C. for anadditional 2 days, and then evaluated for colony growth relative tocontrol. Experimental results are summarized in Table 2. The resultssuggest that Compound A is able to inhibit Escherichia coli.

TABLE 8 Results of in vitro assay for volatile fungicide Rate ofCompound A (mg per disk) Colony Rating 5.00 1 Untreated 3 Not Inoculated0 Colony Rating: 0 = No colonies 1 = Micro colonies not connected 2 =Small colonies with some merging 3 = Large colonies merging together

Example 9

For testing activities against additional fungi pathogens, 12-Well (6.5mL volume per well) microtiter plates are used for the in vitroinhibition assay for volatile antimicrobial compounds. A 3-mL volume offull-strength Potato Dextrose Agar (PDA) is added to each well. Aftercooling, 1 μL of 1×10⁵ per mL Botrytis cinerea, Penicillium expansum,Alternaria alternata, Monilinia fructicola or Glomerella cingulata sporesuspension is spot-pipetted to the center of the agar. Plates areinoculated immediately prior to volatile fungicide treatment. A Whatman#1 filter disk (Cat. No. 1001-0155) is placed, in duplicate, on theunderside of a polyethylene PCR plate sealing film. For determination ofthe minimum inhibitory concentration (MIC), compounds are diluted inacetone, and the appropriate amount of compound is added to the disks ina dose dependent manner to achieve a final headspace concentration of1142.9 to 0.6 mg/L. The acetone is permitted to evaporate for 5 minutes.The headspace around the inoculum is then sealed inside the well by thefilm with the adhering disk containing the fungicide by inverting theplates over the treated disks and sealing to prevent any of the chemicalfrom flaking from the disk and falling onto the inoculated agar. After 3days of storage at 23° C., the cultures are evaluated for percent growthrelative to control based on measurement of fungal colony diameter.Experimental results are summarized in Table 3. The results indicatethat benzoxaborole compounds have excellent in vitro activity againstfive selected fungal pathogens.

TABLE 9 MIC (mg/L, headspace concentration) of numerous benzoxaborolecompounds applied as a volatile treatment against numerous fungalpathogens (Compound 10 is the same as Compound A, and Compound 11 is thesame as Compound B2). Cmpd MIC (mg/L) Structure # BOTRCI PENIEX ALTEALMONIFC GLOMCI

6 2.2 17.9 4.5 8.9 17.9

7 2.2 17.9 8.9 8.9 71.4

8 2.2 35.7 8.9 4.5 71.4

9 2.2 8.9 8.9 8.9 35.7

10 2.2 2.2 <0.6 <0.6 <0.6

11 4.5 17.9 4.5 2.2 35.7

30 2.2 8.9 2.2 2.2 n/a

34 <0.6 2.2 2.2 n/a n/a

200 10.6 68.3 7.3 6.3 n/a

201 3.8 29.5 16.1 8.5 9.3 BOTRCI = Botrytis cinerea PENIEX = Penicilliumexpansum ALTEAL = Alternaria alternata MONIFC = Monilinia fructicolaGLOMCI = Glomerella cingulata

Example 10

12-Well (6.5 mL volume per well) microtiter plates are used for the invitro inhibition assay for volatile antimicrobial compounds. A 3-mLvolume of full-strength Potato Dextrose Agar (PDA) is added to eachwell. After cooling, 1 μL of 1×10⁵ per mL Botrytis cinerea andPenicillium expansum spore suspension is spot-pipetted to the center ofthe agar. Plates are inoculated immediately prior to volatile fungicidetreatment. A Whatman #1 filter disk (Cat. No. 1001-0155) is placed, induplicate, on the underside of a polyethylene PCR plate sealing film.For determination of the minimum inhibitory concentration (MIC),compounds are diluted in acetone, and the appropriate amount of compoundis added to the disks in a dose dependent manner to achieve a finalheadspace concentration of 35.7 to 0.03 mg/L. The acetone is permittedto evaporate for 5 minutes. The headspace around the inoculum is thensealed inside the well by the film with the adhering disk containing thefungicide by inverting the plates over the treated disks and sealing toprevent any of the chemical from flaking from the disk and falling ontothe inoculated agar. After 3 days of storage at 23° C., the cultures areevaluated for percent growth relative to control based on measurement offungal colony diameter. Experimental results are summarized in Table 10.The results indicate that numerous benzoxaborole compounds haveexcellent in vitro activity against two selected fungal pathogens.

TABLE 10 MIC (mg/L) of numerous benzoxaborole compounds applied as avolatile treatment against Botrytis cinerea and Penicillium expansumfungal pathogens. Cmpd MIC (mg/L) Structure # BOTRCI PENIEX

21 1.1 35.7

22 4.5 35.7

38 0.6 8.9

39 0.6 8.9

54 0.6 4.5

55 4.5 >35.7

62 2.2 8.9

63 1.1 17.9

64 1.1 8.9

72 35.7 >35.7

73 35.7 >35.7

74 2.2 35.7

86 0.6 8.9

87 0.6 8.9

105 0.6 4.5

114 17.9 >35.7

115 0.6 8.9

116 1.1 8.9

121 4.5 17.9

122 2.2 17.9

124 4.5 8.9

127 2.2 4.5

129 4.5 8.9

130 1.1 4.5

132 1.1 4.5

133 8.9 35.7

134 17.9 >35.7

135 17.9 >35.7

136 8.9 >35.7

137 0.3 1.1

202 35.7 142.9

203 8.9 142.9

204 8.9 >35.7 BOTRCI = Botrytis cinerea (gray mold) PENIEX = Penicilliumexpansum (blue mold)

Example 11

12-Well (6.5 mL volume per well) microtiter plates are used for the invitro inhibition assay for volatile antimicrobial compounds A and B2against additional fungal pathogens.

A 3-mL volume of full-strength Potato Dextrose Agar (PDA) is added toeach well. After cooling, 1 μL of 1×10⁵ spores per mL of Botrytiscinerea, Penicillium expansum, Alternaria alternata, Glomerellacingulata, Penicillium digitatum, Monilinia fruticola, Aspergillusbrasiliensis, Colletotrichum acutatum, Fusarium sambucinum, Phytophthoracapsici, Geotrichum candidum, Aspergillus niger, Diplodia gossypina orDiaporthe citrii suspension is spotted onto the center of the agar. AWhatman #1 filter disk (Cat. No. 1001-0155) is placed, in duplicate, onthe underside of a polyethylene PCR plate sealing film. Fordetermination of the minimum inhibitory concentration (MIC), testcompounds are diluted in acetone, and the appropriate amount of compoundis added to the disks in a dose dependent manner to achieve a finalheadspace concentration of 35.7 to 0.03 mg/L. The acetone is permittedto evaporate for five minutes. The headspace around the inoculum is thensealed inside the well by the film with the adhering disk containing thefungicide by inverting the plates over the treated disks and sealing toprevent any of the chemical from flaking from the disk and falling ontothe inoculated agar. After 3 days of storage at 23° C., cultures areevaluated for percent growth relative to control. Results shown in Table11 demonstrate the ability of benzoxaborole compounds A and B2 tocontrol the growth of numerous fungal pathogens through volatileactivity.

TABLE 11 MIC (mg/L) of Compounds A and B applied as a volatile againstnumerous fungal pathogens Compound A Compound B2 Pathogens MIC MIC B.cinerea 2.2 4.5 P. expansum 1.1 8.9 M. fruticola 2.2 1.1 A. alternata2.2 2.2 G. cingulata 17.9 35.7 P. digitatum 2.2 4.5 A. brasiliensis 2.20.6 C. acutatum 4.4 8.9 F. sambucinum 1.1 4.5 P. capsici 1.1 n/a G.candidum 8.9 8.9 A. niger 2.2 1.1 M. piriformis 1.1 2.2 D. gossypina 1.14.5 D. citrii 2.2 17.9

Example 12

12-Well (6.5 mL volume per well) microtiter plates are used for the invitro inhibition assay for volatile antimicrobial Compound A againstadditional bacterial pathogens. A 3-mL volume of Nutrient agar is addedto each well and allowed to dry before introducing the pathogen.Escherichia coli, Pectobacterium carotovorum, Xanthomonas axonopodis andSalmonella enterica cell suspensions are adjusted to an optical densityof 0.2 to 0.35, and further diluted 1/10, and 15 μL is pipetted to thecenter of each well and tilted to distribute uniformly. A Whatman #1filter paper (CAT 1001-0155) is placed on the underside of apolyethylene PCR plate sealing film. For determination of minimumbactericidal concentration (MBC), Compound A is diluted in acetone, and50 μL are applied to the disks, in duplicate, in a dose dependent mannerin order to achieve a final headspace concentration of 71.4 to 0.03mg/L. The acetone is permitted to evaporate for 5 minutes. The filmswith the treated disks are then applied over the inoculated plates andsealed. Plates are inverted, and incubated at 23° C. for 48 hours. Afterthe incubation period, the bacteria colonies are dislodged in sterilewater containing tween 80 (0.001%) and the optical density (OD; 600 nm)is determined. Results are summarized in Table 6, where the headspaceconcentration required to control at least 80% of bacterial growth isreported. Compound A shows good antimicrobial activity against numerousbacteria in this in vitro assay.

TABLE 12 Rate (mg/L) of Compound A offering at least 80% control againstbacterial pathogens E. coli P. carotovorum X. axonopodis S. enterica35.7 2.2 4.5 17.9

Example 13

An in vitro assay is used to evaluate the ability of Compound A tovolatilize from different materials and control fungal growth.PTFE-Coated Fiberglass (8577K81), Fiberglass (8816K1), Silica (8799K3),Aramid and Fiberglass blend (8821K4), Vinyl-Coated Polyester (8843K31),Acrylic-Coated Fiberglass (8838K2), Silicone-Coated Fiberglass(87815K1), Aramid (1206T1) (all McMaster-Carr, Santa Fe Springs,Calif.), Polyethylene PCR sealing film, Cellulose (Whatman #1, Cat no.1001-0155), PTFE (Cole Parmer, Cat no. 36229-32), and Category-1cardboard were cut into disks of 15 mm diameter. 12-Well (6.5 mL volumeper well) microtiter plates are used for the in vitro inhibition assayfor volatile antimicrobial compounds. A 3-mL volume of full-strengthPotato Dextrose Agar (PDA) is added to each well. After cooling, 1 μL of1×10⁵ per mL Botrytis cinerea spore suspension is spot-pipetted to thecentre of the agar. Plates are inoculated immediately prior to volatilefungicide treatment.

TABLE 13 Effects of different materials on the volatile release ofCompound A and the subsequent in vitro inhibition (MIC) of Botrytiscinerea. Material MIC (mg/L) Polyethylene PCR Film 0.28 PTFE- CoatedFiberglass 0.56 Fiberglass 0.56 Cellulose 0.56 Silica 0.56 Aramid andFiberglass 0.56 Vinyl-Coated Polyester 0.56 Acrylic-Coated Fiberglass0.56 Silicone- Coated Fiberglass 0.56 PTFE 1.1 Cardboard 2.2 Aramid 2.2

The various materials are placed, in duplicate, on the underside of apolyethylene PCR plate sealing film. For determination of the minimuminhibitory concentration (MIC), compounds are diluted in acetone, andthe appropriate amount of compound is added to the materials in a dosedependent manner to achieve a final headspace concentration of 35.7 to0.03 mg/L. The acetone is permitted to evaporate for five minutes. Theheadspace around the Botrytis cinerea inoculum is then sealed inside thewell by the film with the adhering disk of material containing thefungicide. Plates are inverted, placed over the treated disks and sealedto prevent any of the chemical from flaking from the disk and fallingonto the inoculated agar. After three days of storage at 23° C., thecultures are evaluated for percent growth relative to control based onmeasurement of fungal colony diameter. Experimental results aresummarized in Table 7. The results indicate that Compound A canvolatilize from numerous materials to inhibit the in vitro growth ofBotrytis cinerea with similar levels of control.

Example 14

An in vitro assay is used to evaluate the ability of compound A tovolatilize from different materials and control fungal growth. Cardboardbox (Category 1), PET plastic (Polyethylene terepthalate—PET) andPolyethylene are used. The materials are cut into equal dimensions(10×19 cm²) and placed inside a 36-L acrylic desiccator cabinet (FisherScientific, cat no. 08-642-23C) in duplicate.

TABLE 14 Effects of different materials on the volatile release ofCompound A and the subsequent in vitro inhibition of Botrytis cinereaMaterial Incidence (%) Rate (mg/L) Clamshell Cardboard Polyethylene 0.34.1 9.3 4.9 0.06 100.0 91.7 86.7 0.012 100.0 100.0 99.0

Compound A is dissolved in acetone and 100 μL of the solution pipettedinto a glass tube. The acetone is allowed to evaporate for 1 minute at60° C. Compound A is then introduced as a gas into the cabinets by asublimation device (copper tube heated to 180° C. with fan flow at 0.5L/min) to achieve a final headspace concentration of 0.3, 0.06 and 0.012mg/L). The chambers are then incubated at 23° C. for 24 hours, thentreated materials are carefully removed and placed inside a clean 10.8cup SnapWare airtight container (Model #109842) containing a 10-cmdiameter Petri dish with PDA and inoculated with 1 μL of 1×10 spores/mLof B. cinerea. The containers are then tightly sealed for 3 days at 23°C. After 3 days of storage, cultures are evaluated for percent growthrelative to control. Table 14 demonstrates the ability of benzoxaborolecompounds A to control the growth of B. cinerea through volatileactivity.

Example 15

3.20 g of 5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (21.2 mmol)and 3.20 g of ethylene glycol (51.6 mmol) are heated in 40 g of toluene.The toluene water azeotrope is distilled out of the system until thehead temperature reached 110° C. The toluene is removed via rotaryevaporator and the excess ethylene glycol is removed by kugelrohrdistillation at about 20 torr and 100° C. bath temperature.Recrystallization from toluene generates 2.95 g of white crystals, mp145-149° C. Proton nmr shows spectra and integration consistent with thetwo to one product below:

Example 16

Preparation of Sample 2

3.00 g of 1,3-dihydro-1-hydroxy-2,1-benzoxaborole (22.4 mmol) and 3.00 gof ethylene glycol (46.9 mmol) are heated in 40 g of toluene. Thetoluene water azeotrope is distilled out of the system until the headtemperature reached 110° C. The toluene is removed via rotary evaporatorand the excess ethylene glycol is removed by kugelrohr distillation atabout 20 torr and 100° C. bath temperature. Recrystallization fromtoluene generates 2.49 g of white crystals, mp 118-120.5° C. Proton NMRshows spectra and integration consistent with the two to one product.

Example 17

Preparation of Sample 3

3.17 g of 5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (21.0 mmol)and 3.22 g of pinacol (27.3 mmol) are heated in 40 g of toluene. Thetoluene water azeotrope is distilled out of the system until the headtemperature reached 110° C. The toluene is removed via rotary evaporatorand the excess pinacol is removed by kugelrohr distillation at about 20torr and 120° C. bath temperature. Recrystallization from hexanegenerates 3.21 g of white crystals, mp 81-89° C. Proton NMR showsspectra and integration consistent with the two to one product.

Example 18

Preparation of Sample 4

3.0 g of 5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (19.9 mmol)and 2.5 g of 1,2-propanediol (propylene glycol; 32.9 mmol) are heated in40 g of toluene. The toluene water azeotrope is distilled out of thesystem until the head temperature reached 110° C. The toluene is removedvia rotary evaporator and the excess propylene glycol is removed bykugelrohr distillation at about 20 torr and 110° C. bath temperature.Recrystallization from hexane generates 3.49 g of white crystals, mp65.5-68.5° C. Proton NMR shows spectra and integration consistent withthe two to one product.

Example 19

In Vitro Analysis

12-well (6.5 ml volume per well) microtiter plates are used for the invitro inhibition assay for volatile antimicrobial compounds. A 3-mlvolume of full-strength Potato Dextrose Agar (PDA) is added to eachwell. After cooling, 1 μL of 1×10⁵ spores per ml Botrytis cinerea (ATCC#204446) spore suspension is spot pipetted to the agar in the centre ofthe well.

TABLE 15 Antimicrobial activities of Samples 1-4 (50 μl/disk) MIC mg/lBotrytis Penicillium Alternaria Monilinia Glomerella ID cinerea expansumalternata fructicola cingulata Sample 1 <0.6 8.9 22 — — Sample 2 <0.68.9 8.9 35.7 142.9 Sample 3 <0.6 4.5 2.2 — — Sample 4 <0.6 8.9 1.1 — —

Whatman #1 filter disks (1.5 cm; Cat. No. 1001-0155) are placed on theunderside of a polyethylene PCR plate sealing film. For determination ofthe minimum inhibitory concentration (MIC), test compounds are dilutedin acetone, in duplicate, and 50 μl of the compound solution is added todisks at concentrations that can vary from 0.001 mg/l to 1142.9 mg/l.

TABLE 16 Antimicrobial activities of Samples 1-4 (repeat test; 50μl/disk) MIC mg/l Botrytis Penicillium Alternaria Monilinia GlomerellaID cinerea expansum alternata fructicola cingulata Sample 1 0.6 8.9 >2.22.2 — Sample 2 2.2 8.9 — — — Sample 3 1.1 8.9 >2.2 1.1 — Sample 4 0.68.9 >2.2 1.1 —

The acetone is permitted to evaporate for 5 minutes. The headspacearound the Botrytis cinerea inoculum is then sealed inside the well bythe film with the adhering disk containing the fungicide. Plates areinverted to prevent any possibility of the chemical from flaking fromthe disk and falling onto the inoculated agar. After 3 days ofincubation at 23° C., cultures are evaluated for percent growth relativeto control and determination of MIC. Samples 1-4 show good antimicrobialactivity against Botrytis cinerea and/or other pathogens in this invitro analysis. Minimum inhibitory concentrations (MIC) are shown inTables 9 and 10 for results from two separate tests.

Example 20

Antimicrobial Activity Against Bacteria

12-well (6.5 ml volume per well) microtiter plates are used for the invitro inhibition assay for volatile antimicrobial compounds. A 3-mlvolume of full-strength LB Agar is added to each well. After cooling, 15μL of Escherichia coli (ATCC #25922) adjusted to an optical density of0.02 to 0.035, and further diluted 1/10 is pipetted to the centre of theagar. The plate is tilted to distribute bacteria uniformly. Whatman #1filter disks (1.5 cm; Cat. No. 1001-0155) are placed on the underside ofa polyethylene PCR plate sealing film. For determination of the minimuminhibitory concentration (MIC), test compounds are diluted in acetone,in duplicate, and 50 μl of compound is added to disks at concentrationsthat can vary from 0.015 to 35.7 mg/l. The acetone is permitted toevaporate for 5 minutes. The headspace around the Escherichia coliinoculum is then sealed inside the well by the film with the adheringdisk containing the fungicide. Plates are inverted, placed over thetreated disks and sealed to prevent any of the chemical from flakingfrom the disk and falling onto the inoculated agar. After 2 days ofincubation at 23° C., cultures were evaluated for colony growth relativeto control. Samples 1-4 show good antimicrobial activity againstEscherichia coli in this in vitro analysis.

Example 21

In order to demonstrate unexpected volatility of compound A(1-hydroxy-5-fluoro-1, 3-dihydro-2, 1-benzoxaborole) and a new method toapply the volatile compound A, another in vivo assay is developed tocontrol Botrytis cinerea on strawberry. Eight strawberries (perrepetition, in triplicate) are placed in an industry standard 1-lb PETclamshell with the stem-end facing down. A fresh wound on the upwardsfacing tip of the fruit is then inoculated with 20 μL of 1×10⁵ sporesper mL suspension of B. cinerea.

Two identically prepared clamshells per repetition and treatment arethen placed at the bottom of a 36-L acrylic desiccator cabinet (FisherScientific, No. 08-642-23C), and pre-cooled for 2 hours at 1° C. priorto treatment application. Compound A is then mixed with acetone and 100μL of the mixture is pipetted into a small glass tube. This tube is thenplaced inside a pre-heated sublimation device (0.5″ OD by 6″ longthermostatically heated copper tube mounted to a 0.5 L/min low flow fan)set at 60° C. for 1-minute to allow the acetone to evaporate. Compound Ais then introduced into the cabinet containing the clamshell by usingthe sublimation device set at 180° C. to achieve a final headspaceconcentration of 0.1 mg/L and equilibrated at 1° C. for 0.5 or 1 hour.

TABLE 17 Valatile application of Compound A to control B. cinereainfection Clamshell Treatment Time Disease Severity (0 to 4) Condition(hour) Day 1 Day 3 Day 5 Untreated 0 1.3 3.5 4.0 Treated 0.5 0.0 0.1 2.31 0.0 0.0 0.4 Untreated (fruit 0.5 0.0 2.7 3.4 transfer) 1 0.0 0.1 1.6

After incubation, both clamshells are removed from the treatmentchamber. One clamshell is undisturbed while the fruit from the secondclamshell are immediately transferred into a new untreated clamshell.All clamshells are then held at 1° C. for 5 days and then evaluatedduring an additional 5 days at 21° C. During the 5 days at 21° C., thefruits are evaluated for gray mold severity (scale 0 to 4, with ≤1indicating marketable fruit and 4 indicating ≥50% of fruit surfacecovered by pathogen). The results from Table 11 demonstrate theunexpected volatility of Compound A applied to clamshells and itsability to control B. cinerea development on strawberry throughout the 5days of simulated marketing at 21° C. The treated clamshell producesmarketable fruit up to 3 days with 0.5 hour treatment (0.1), whereasfruits in new clamshells are unmarketable (2.7). Similarly, the treatedclamshell produces marketable fruit up to 5 days with 1 hour treatment(0.4), whereas fruits in new clamshells are unmarketable (1.6). Thus,treatments where berries remain in the treated clamshells have the bestlevel of control due to the compound further volatilizing over time offof the clamshell surface.

Furthermore, berries that are placed into a new clamshell still benefitfrom the initial volatile treatment and demonstrated better control ofBotrytis cinerea than untreated fruit, but the control is less than thetreated clamshell since there is no longer any new exposure to thevolatile substance off of the treated clamshell surface. Therefore, theresults from this study provide evidence that a volatile application ofCompound A provides control of fungal pathogen growth (untreated fruittransfer) and that Compound A deposited on clamshell surfaces willsubsequently volatilize during 5 days at 21° C., providing additionaluseful control of Botrytis cinerea growth (treated).

Example 21

In order to demonstrate unexpected volatility of Compound A, another invivo assay is developed to evaluate blue mold (Penicillium expansum)control on apple. Two apples are placed in a clamshell, and three freshwounds are made near the equatorial region of each fruit. Each fruitwound is then inoculated with 20 μL of 1×10⁶ spores per mL ofPenicillium expansum suspension. The inoculum is allowed to dry for twohours prior to treatment application as a volatile or contact.

TABLE 18 Comparison of volatile and contact fungicidal activity ofCompound A to control Penicillim expansum infection Treatment rateBrowning Rot (diameter; mm) Assay (mg/L) Day 0 Day 2 Day 4 Day 7 Contact0 0.0 6.4 15.7 29.6 2 0.0 3.7 15.5 23.3 10 0.0 2.2 8.0 20.2 50 0.0 0.75.7 15.0 250 0.0 0.0 4.3 11.8 Volatile 0 0.0 6.6 16.1 30.5 0.02 0.0 0.92.8 6.9 0.1 0.0 0.0 0.3 2.0 0.5 0.0 0.0 0.0 0.4 2.5 0.0 0.0 0.0 0.0

Volatile Assay: Clamshells are then placed at the bottom of a 36-Lacrylic desiccator cabinet (Fisher Scientific, No. 08-642-23C). CompoundA is mixed with acetone and 250 μL of the mixture is pipetted into asmall glass tube. This tube is then placed inside a pre-heatedsublimation device (0.5″ OD by 6″ long thermostatically heated coppertube mounted to a 0.5 L/min low flow fan) set at 60° C. for 1 minute toallow the acetone to evaporate. Compound A is then introduced into thecabinets containing the clamshells by using the sublimation device setat 180° C. to achieve a final headspace concentration of 2.5, 0.5, 0.1or 0.02 mg/L. The chambers are then incubated at 1° C. for 5 days. Afterincubation, fruits are evaluated by measuring the diameter (mm) of rotdevelopment (browning) up to 7 days at 21° C.

Contact Assay: Compound A is dissolved in 85% methanol to achieve afinal concentration of 250, 50, 10, or 2 mg/L. A 250 mL solution of eachconcentration is used to dip two inoculated apples, one minute perapple, performed in triplicate per rate. The dipped fruits are thenplaced back into the clamshells, which are then placed in a secondarycontainer and incubated at 1° C. for 5 days. After incubation, fruitsare evaluated for diameter (mm) of rot development (browning) up to 7days at 21° C. Table 12 demonstrates the unexpected volatility ofCompound A to control Penicillium expansum on apples during storage evenwhen applied at 100× lower rate (v/v) than as a contact.

Example 23

In order to demonstrate unexpected volatility of Compound A, another invivo assay is developed to evaluate gray mold (Botrytis cinerea) controlon strawberry. Eight strawberries (per repetition, in triplicate) areplaced in an industry standard 1-lb PET clamshell with the stem-endfacing down. A fresh wound on the upwards facing tip of the fruit isthen inoculated with 20 μL of 1×10⁵ spores per mL suspension of B.cinerea. The inoculum is allowed to dry for two hours prior to treatmentapplication as a volatile or contact.

TABLE 19 Comparison of volatile and contact fungicidal activity ofCompound A to control Botrytis cinerea infection Disease Severity (0 to4) Assay Treatment rate (mg/L) Day 0 Day 1 Day 2 Day 3 Day 4 Contact 00.0 0.0 0.8 1.1 2.0 2 0.0 0.0 0.6 1.1 1.9 10 0.0 0.0 0.0 0.3 1.0 50 0.00.0 0.0 0.0 0.0 250 0.0 0.0 0.0 0.0 0.0 Volatile 0 0.0 0.1 0.9 1.4 2.30.02 0.0 0.0 0.1 0.2 0.7 0.1 0.0 0.0 0.0 0.0 0.0 0.5 0.0 0.0 0.0 0.0 0.02.5 0.0 0.0 0.0 0.0 0.0

Volatile Assay: Clamshells are then placed at the bottom of a 36-Lacrylic desiccator cabinet (Fisher Scientific, No. 08-642-23C). CompoundA is mixed acetone and 250 μL of the mixture is pipetted into a smallglass tube. This tube is then placed inside a pre-heated sublimationdevice (0.5″ OD by 6″ long thermostatically heated copper tube mountedto a 0.5 L/min low flow fan) set at 60° C. for 1-minute to allow theacetone to evaporate. Compound A is then introduced into the cabinetscontaining the clamshells by using the sublimation device set at 180° C.to achieve a final headspace concentration of 2.5, 0.5, 0.1 or 0.02mg/L. The chambers are then incubated at 1° C. for 5 days. Afterincubation, fruits are evaluated for disease (scale 0 to 4, with ≤1indicating marketable fruit and 4 indicating ≥50% of fruit surfacecovered by pathogen) up to 4 days at 21° C.

Contact Assay: Compound A is dissolved in 85% methanol to achieve afinal concentration of 250, 50, 10, or 2 mg/L. A 250 mL solution of eachconcentration is used to dip eight inoculated strawberry fruit forone-minute, performed in triplicate per rate. The dipped fruits are thenplaced back into the clamshells, which are then placed in a secondarycontainer and incubated at 1° C. for 5 days. After incubation, fruitsare evaluated for disease severity (scale 0 to 4, with ≤1 indicatingmarketable fruit and 4 indicating ≥50% of fruit surface covered bypathogen) up to 4 days at 21° C. Table 19 demonstrates the unexpectedvolatility of Compound A to control Botrytis cinerea on strawberriesduring storage even when applied at 100× lower rate (v/v) than as acontact.

Example 24

An in vitro assay was performed comparing the volatile and contactactivity of various benzoxaborole compounds to demonstrate the activityof compound 10 relative to other similar structures from the chemicalclass.

Volatile assay: 12-well (6.5 mL volume per well) microtiter plates areused. A 3-mL volume of half strength PDA is added to each well. Aftercooling, 1 μL of 1×10⁵ spores per mL of Botrytis cinerea or Penicilliumexpansum suspension is spotted to the center of the agar. A Whatman #1filter disk (Cat. No. 1001-0155) is placed, in duplicate, on theunderside of a polyethylene PCR plate sealing film. Test compounds aremixed with acetone and the mixtures are added to disks in a dosedependent manner to achieve a final headspace concentration of 35.7 to0.03 mg/L. The acetone is permitted to evaporate for 5 minutes. Theheadspace around the inoculum is then sealed inside the well by the filmwith the adhering disk containing the fungicide. Plates are inverted,placed over the treated disks, and sealed to prevent any of the chemicalfrom flaking from the disk and falling onto the inoculated agar. After 3days of incubation at 23° C., cultures are evaluated for percent growthrelative to the acetone only control.

TABLE 20 Comparison of contact and volatile activity of selectedbenzoxaborole compounds Compound Contact MIC (mg/L) Volatile MIC (mg/L)ID B. cinerea P. expansum B. cinerea P. expansum  6 2.0 2.0 4.5 17.9 10* <0.08 0.4 0.3 2.2  11* 10 2.0 4.5 17.9  31 0.4 2.0 0.6 8.9  33 0.42.0 0.6 8.9  34 2.0 0.4 0.6 2.2 121 10.0 10.0 4.5 17.9 124 2.0 2.0 4.58.9 130 2.0 0.4 1.1 4.5 132 2.0 2.0 1.1 4.5 135 2.0 2.0 17.9 >35.7*Compound 10 is identical to Compound A; Compound 11 is identical toCompound B2

Contact assay: 6-well (16.5 mL volume per well) microtiter plates areused for an in vitro inhibition assay. Half-strength Potato DextroseAgar (PDA) is amended with a mixture of one of the test compounds withacetone or methanol to a final concentration of 50 to 0.08 mg/L. A7.5-mL volume of the amended media is added to each well of themicrotiter plate. After drying, 1 μL of 1×10⁵ spores per mL of B.cinerea or P. expansum suspension is spotted to the center of the agar.The plates are sealed with a clear film and incubated for 3 days at 23°C. After incubation, plates are evaluate for percent growth relative toacetone only control. Results are reported as the minimum inhibitoryconcentration (MIC) required for 100% control of pathogen growth. Table20 shows the MIC results of numerous benzoxaboroles assayed for bothcontact and volatile activity. Results demonstrate that numerousstructures in the benzoxaborole class of compounds have both contact andvolatile activity.

TABLE 21 Compounds used in this example Compound ID Benzoxaborolestructure  6

 10*

 11*

 31

 33

 34

121

124

130

132

135

*Compound 10 is identical to Compound A; Compound 11 is identical toCompound B2 Compound 33 is identical to Compound B

Example 25

In order to demonstrate unexpected volatility of Compound A, another invivo assay is developed to evaluate blue mold (Penicillium expansum)control on apple and pear, as well as green mold (Penicillium digitatum)control on orange. Two apples, pears or oranges are placed in aclamshell, and three fresh wounds are made near the equatorial region ofeach fruit. Each fruit wound is then inoculated with 20 μL of 1×10⁶spores per mL of Penicillium expansum or digitatum suspension,respectively. The inoculum is allowed to dry for two hours prior totreatment application as a volatile or contact.

TABLE 22 Comparison of Compound A with other fungicides in volatile andcontact assays Apple Pear Orange Test Browning Sporulation BrowningSporulation Sporulation Assay Compound (mm) (mm) (mm) (mm) (mm) VolatileControl 11.1 2.2 14.9 4.0 44.9 (acetone only) Compound A 0.4 0.0 5.7 0.00.0 Control 9.7 2.3 13.4 4.4 39.3 (ethanol only) Boscalid 10.3 2.4 14.33.6 30.3 Fludioxonil 11.2 3.1 12.8 2.8 40.8 Imazalil 11.1 3.0 14.3 3.441.4 Pyrimethanil 11.3 2.6 14.0 6.5 22.3 Thiabendazole 8.4 2.0 12.93.0 >50 Contact Control 10.3 2.6 15.1 5.4 >50 (5% PG only) Compound A9.9 2.3 18.9 8.4 >50 Control 9.2 5.1 8.9 1.9 9.7 (ethanol only) Boscalid7.0 0.8 8.6 2.3 >50 Fludioxonil 2.9 0.0 3.3 0.0 8.3 Imazalil 6.9 0.9 7.91.0 0.0 Pyrimethanil 8.1 2.4 8.9 5.2 0.0 Thiabendazole 8.2 1.9 9.1 5.40.0

Volatile Assay: Clamshells are then placed at the bottom of a 2.55-LSnapWare airtight container (Model #109842). An appropriate amount ofCompound A (dissolved in acetone), Boscalid, Fludioxinil, Imazalil,Pyrimethanil or Thiabendazole (methanol) is solubilized to achieve atreatment rate of 50 mg/L. (Compound A is not soluble in methanol atroom temperature). The solutions are pipetted into Whatman filter disksmounted to the inside lid of the container. The chambers are thenincubated at 1° C. for 5 days, removed to 21° C., and evaluated on day 3by determining the diameter (mm) of rot development (browning) orsporulation.

Contact Assay: Compound A is dissolved in 5% propylene glycol, whereasall other actives are dissolved in 85% methanol at a rate to achieve afinal concentration of 250, 50, 10, or 2 mg/L (Compound A is not solublein methanol at room temperature). A 250 mL solution of eachconcentration is used to dip two inoculated fruits, one minute perfruit, performed in triplicate per rate. The dipped fruits are thenplaced back into the clamshells and then into the SnapWare container andincubated at 1° C. for 5 days. The containers are then incubated at 1°C. for 5 days, removed to 21° C., and evaluated on day 3 by determiningthe diameter (mm) of rot development (browning) or sporulation. Table 21demonstrates the unexpected volatility of Compound A to controlPenicillium expansum on apples and pears, as well as Penicilliumdigitatum on oranges. Volatile application of Compound A results inexcellent inhibition of browning and sporulation, whereas all otheractive ingredients result in no or little inhibition. However, contactapplication of Compound A does not provide good inhibition of browningand sporulation as compared to other fungicides, demonstrating that thevolatile application is important for the fungicidal activity ofCompound A.

Example 26

In order to demonstrate the volatile activity of Compound A and Compound31 relative to commercially registered fungicides, an in vitro assay isperformed comparing the volatile and contact activity of the activeingredients.

Contact Assay: 12-well (6.5 mL volume per well) microtiter plates areused for the in vitro inhibition assay for Compounds A and 31, andcompared to other registered fungicides (5-fluorocytosine, AmphotericinB, Caspofungin diacetate, Fluconazole and Itraconazole). Half-strengthPotato Dextrose Agar (PDA) is amended with a mixture of one of the testcompounds in acetone or methanol to a final concentration of 50, 10, 2,0.4 or 0.08 mg/L. A 3-mL volume of the amended media is added to eachwell of the microtiter plate. After drying, a mycelial plug (5 mmdiameter) is aseptically obtained from actively growing cultures ofEpidermophyton floccus, Trichophyton rubrum, or Trichophytonmentagrophytes and placed at the center of the plate with the mycelialside in contact with the agar. The plates are sealed with a clear filmand incubated inverted for 5 days at 28° C. After incubation, culturesare evaluated (mm diameter growth) for percent growth relative tocontrol with results expressed as minimum inhibitory concentration (MIC)required to control 100% of pathogen growth.

TABLE 23 Comparison of Compounds A and 31 together with other fungicideto control selected fungal pathogens MIC (mg/L) Epider- TrichophytonTest mophyton Trichophyton mentagr- Assay Compound floccus rubrumophytes Contact Compound A 2.0 2.0 2.0 Compound 31 n.d. 10.0 2.05-Fluorocytosine >50 >50 >50 Amphotericin B 50.0 >50 >50 Caspofungin 2.010.0 50.0 Diacetate Fluconazole 10.0 50.0 >50 Itraconazole >50 >50 >50Volatile Compound A 2.0 2.0 2.0 Compound 31 n.d. 2.0 2.05-Fluorocytosine >50 >50 >50 Amphotericin B >50 >50 >50Caspofungin >50 >50 >50 Diacetate Fluconazole >50 >50 >50Itraconazole >50 >50 >50 n.d. = not determined.

Volatile assay: 6-well (16.5 mL volume per well) microtiter plates areused in an in vitro inhibition assay for Compounds A and 31, andcompared to other registered fungicides (5-fluorocytosine, AmphotericinB, aspofungin diacetate, Fluconazole and Itraconazole). A 7.5-mL volumeof half strength PDA is added to each well. After drying, a mycelialplug (5 mm diameter) is aseptically obtained from actively growingcultures of Epidermophyton floccus, Trichophyton rubrum, or Trichophytonmentagrophytes and placed at the center of the plate with the mycelialside in contact with the agar. A Whatman #1 filter disk (Cat. No.1001-325) is placed, in duplicate, on the underside of a polyethylenePCR plate sealing film. For determination of unexpected volatility, testcompounds are mixed with acetone or methanol, and then added to disks ina dose dependent manner to achieve a final headspace concentration of50, 10, 2, 0.4 or 0.08 mg/L. The acetone/methanol is permitted toevaporate for 5 minutes. The headspace around the inoculum is thensealed inside the well by the film with the adhering disk containing thefungicide and incubated inverted for 5 days at 28° C. After incubation,cultures are evaluated for percent growth relative to control withresults expressed as minimum inhibitory concentration (MIC) required tocontrol 100% of pathogen growth.

Volatile application of benzoxaboroles of Compounds A and 31 showsignificant fungicidal activities. Table 23 demonstrates the unexpectedvolatile activity of Compounds A and 31 with a minimum inhibitoryconcentration (MIC) of 2 mg/L for both Compound A and 31. In comparison,none of the commercial fungicide standards demonstrated any significantvolatile activity where little or no fungicidal activity after volatileapplications.

Example 27

In order to demonstrate the volatile activity of Compound 10 (i.e.,Compound A) on fungal pathogen species causing human yeast infection, anin vitro assay to measure the rate of growth inhibition was performed.More specifically, growth inhibition of yeast infection fungal speciesC. albicans and C. krusei (or I. orientalis) by volatile treatment ofCompound 10 was assessed via a volatile assay.

Volatile assay: 6-well (16.5 mL volume per well) microtiter plates areused in an in vitro inhibition assay for Compound 10 (i.e., Compound A).A 7.5-mL volume of half strength PDA is added to each well. Afterdrying, a mycelial plug (5 mm diameter) is aseptically obtained fromactively growing cultures of C. albicans and C. krusei (or I.orientalis) and placed at the center of the plate with the mycelial sidein contact with the agar. A Whatman #1 filter disk (Cat. No. 1001-325)is placed, in duplicate, on the underside of a polyethylene PCR platesealing film. For determination of unexpected volatility, test Compound10 is mixed with acetone or methanol, and then added to disks in a dosedependent manner to achieve a final headspace concentration of 35.7,17.9, 8.9, 4.5, 2.2, 1.1, 0.6, 0.3, 0.1, 0.07, 0.035, and 0.017 mg/L(see Table 24). The acetone/methanol is permitted to evaporate for 5minutes. The headspace around the inoculum is then sealed inside thewell by the film with the adhering disk containing the fungicide andincubated inverted for 5 days at 28° C. After incubation, cultures areevaluated for percent growth relative to control with results expressedas minimum inhibitory concentration (MIC) required to control 100% ofpathogen growth.

TABLE 24 MIC (mg/L) of Compound 10 (i.e., Compound A) applied as avolatile against Candida albicans and Issatchenkia orientalis GrowthInhibition (%) MIC mg/L C. albicans C. krusei/I. orientalis 35.7 100.0100.0 17.9 100.0 30.0 8.9 100.0 30.5 4.5 100.0 22.0 2.2 100.0 18.2 1.140.2 −0.2 0.6 22.2 −5.2 0.3 18.5 1.2 0.1 11.9 0.4 0.07 7.9 0.5 0.035 1.6−1.4 0.017 2.0 −1.3

Volatile application of benzoxaboroles of Compound 10 (i.e., Compound A)shows significant fungicidal activities against yeast infectionpathogens, C. albicans and C. krusei. For example, Table 24 demonstratesthe unexpected volatile activity of Compound 10 (i.e., Compound A) witha minimum inhibitory concentration (MIC) of 2.2 mg/L for C. albicans anda minimum inhibitory concentration (MIC) of 35.7 mg/L for C. krusei.

Example 28

An in vivo assay is used to evaluate the ability of Compound A(1-hydroxy-5-fluoro-1, 3-dihydro-2, 1-benzoxaborole) to control fungalgrowth of seeds.

TABLE 25 Effect of a 10 mg/L headspace treatment of Compound A incontrolling Aspergillus brasiliensis growth on grains. Fungal growth onPDA (mm) Grains Compound A Control-Acetone Control-No Acetone Barley 012.8 21.7 Corn Dry 0 10.1 22.8 Millet 0 7.2 19.1 Rice 0 7.5 21.6 Rye 08.4 21 Wheat 0 8.1 22.4

Grains consisting of corn, wheat, rice, rye, millet and barley aresurface sterilized with 0.825% NaOCl for 1 minute and rinsed thrice withsterile distilled water. The grains are inoculated by soaking them in a1×10⁶ spores/mL suspension of Aspergillus brasiliensis for 1 minute. Theexcess inoculum is blotted out with a sterile paper towel before platingfive seeds in a Petri plate containing 25 mL of PDA. For determinationof efficacy, Compound A is diluted in acetone and added to 42.5 mmWhatman #1 filter disks (Cat. No. 1001-042) attached to the inner sideof the lid in a dose dependent manner to achieve a final headspaceconcentration of 0.4, 2, or 10 mg/L. The acetone is permitted toevaporate for five minutes before closing plate and sealing it withparafilm. The plates are incubated at 23° C. for three days. Afterstorage, the grains are evaluated for mycelial colony diameter (mm),with results summarized in Table 25. Results demonstrate 100% control ofAspergillus brasiliensis in this in vivo analysis.

The preceding description enables others skilled in the art to utilizethe technology in various embodiments and with various modifications asare suited to the particular use contemplated. In accordance with theprovisions of the patent statutes, the principles and modes of operationof this disclosure have been explained and illustrated in exemplaryembodiments. Accordingly, the present invention is not limited to theparticular embodiments described and/or exemplified herein.

It is intended that the scope of disclosure of the present technology bedefined by the following claims. However, it must be understood thatthis disclosure may be practiced otherwise than is specificallyexplained and illustrated without departing from its spirit or scope. Itshould be understood by those skilled in the art that variousalternatives to the embodiments described herein may be employed inpracticing the claims without departing from the spirit and scope asdefined in the following claims.

The scope of this disclosure should be determined, not only withreference to the above description, but should instead be determinedwith reference to the appended claims, along with the full scope ofequivalents to which such claims are entitled. It is anticipated andintended that future developments will occur in the arts discussedherein, and that the disclosed compositions and methods will beincorporated into such future examples.

Furthermore, all terms used in the claims are intended to be given theirbroadest reasonable constructions and their ordinary meanings asunderstood by those skilled in the art unless an explicit indication tothe contrary is made herein. In particular, use of the singular articlessuch as “a,” “the,” “said,” etc. should be read to recite one or more ofthe indicated elements unless a claim recites an explicit limitation tothe contrary. It is intended that the following claims define the scopeof the disclosure and that the technology within the scope of theseclaims and their equivalents be covered thereby. In sum, it should beunderstood that the disclosure is capable of modification and variationand is limited only by the following claims.

What is claimed is:
 1. A method of treating a food product with anantimicrobial agent, the method comprising: administering abenzoxaborole treatment directly to one or more surfaces of a foodpackaging material, wherein the benzoxaborole treatment comprises one ormore benzoxaborole compounds, drying the one or more surfaces of thefood packaging material, vaporizing the benzoxaborole compound from theone or more surfaces of the food packaging material to treat the foodproduct located therein, wherein the benzoxaborole is of formula (IV),or a salt thereof:

wherein A and D together with the carbon atoms to which they areattached form a 5-, 6-, or 7-membered fused ring which may besubstituted by C₁-C₆-alkyl, C₁-C₆-alkoxy, hydroxy, halogen, nitro,nitrile, amino, amino substituted by one or more C₁-C₆-alkyl groups,carboxy, acyl, aryloxy, carbonamido, carbonamido substituted byC₁-C₆-alkyl, sulfonamido or trifluoromethyl or the fused ring may linktwo oxaborole rings; X is a group —CR⁷R⁸ wherein R⁷ and R⁸ are eachindependently hydrogen, C₁-C₆-alkyl, nitrile, nitro, aryl, arylalkyl orR⁷ and R⁸ together with the carbon atom to which they are attached forman alicyclic ring; and R⁶ is hydrogen, C₁-C₁₈-alkyl, C₁-C₁₈-alkylsubstituted by C₁-C₆-alkoxy, C₁-C₆-alkylthio, hydroxy, amino, aminosubstituted by C₁-C₁₈-alkyl, carboxy, aryl, aryloxy, carbonamido,carbonamido substituted by C₁-C₆-alkyl, aryl or arylalkyl, arylalkyl,aryl, heteroaryl, cycloalkyl, C₁-C₁₈-alkyleneamino, C₁-C₁₈-alkyleneaminosubstituted by phenyl, C₁-C₆-alkoxy or C₁-C₆-alkylthio, carbonylalkyleneamino or a radical of formula (V):

wherein A, D and X are as defined herein before.
 2. The method of claim1, wherein the food product is selected from the group consisting of astrawberry, a raspberry, a blackberry, and a blueberry.
 3. The method ofclaim 1, wherein the benzoxaborole compound is has the structure

or a salt thereof.
 4. The method of claim 1, wherein administering thebenzoxaborole treatment to food packaging material further comprisesembedding the benzoxaborole compound into the food packaging material,impregnating the food packaging material with the benzoxaborolecompound, or coating the food packaging material with the benzoxaborolecompound.
 5. The method of claim 1, wherein the food packaging materialis a chamber.
 6. A method of treating a food product with anantimicrobial agent, the method comprising: administering abenzoxaborole treatment directly to one or more surfaces of a foodpackaging material, wherein the benzoxaborole treatment comprises one ormore benzoxaborole compounds, drying the one or more surfaces of thefood packaging material, vaporizing the benzoxaborole compound from theone or more surfaces of the food packaging material to treat the foodproduct located therein, wherein the benzoxaborole has the structureformula (A):R^(A)-L^(A)-G-L^(B)-R^(B)  (A), wherein each of R^(A) and R^(B) isindependently a of R^(A) and R^(B) is of formula (E):

wherein each R⁶ is independently hydrogen, alkyl, alkene, alkyne,haloalkyl, haloalkene, haloalkyne, alkoxy, alkeneoxy, haloalkoxy, aryl,heteroaryl, arylalkyl, arylalkene, arylalkyne, heteroarylalkyl,heteroarylalkene, heteroarylalkyne, halogen, hydroxyl, nitrile, amine,ester, carboxylic acid, ketone, alcohol, sulfide, sulfoxide, sulfone,sulfoximine, sulfilimine, sulfonamide, sulfate, sulfonate, nitroalkyl,amide, oxime, imine, hydroxylamine, hydrazine, hydrazone, carbamate,thiocarbamate, urea, thiourea, carbonate, aryloxy, or heteroaryloxy;n=1, 2, 3, or 4; B is boron; X²═(CR⁶ ₂)_(m) where m=1 or 2; each ofL^(A) and L^(B) is independently —O— or

each of R and R′ is independently hydrogen, unsubstituted or substitutedC₁₋₁₈-alkyl, arylalkyl, aryl, or heterocyclic moiety; and G is asubstituted or unsubstituted C₁₋₁₈-alkylene, arylalkylene, arylene, orheterocyclic moiety; and acceptable salts thereof.
 7. The method ofclaim 6, wherein the food product is selected from the group consistingof a strawberry, a raspberry, a blackberry, and a blueberry.
 8. Themethod of claim 6, wherein the benzoxaborole has the structure

or a salt thereof.
 9. The method of claim 6, wherein administering thebenzoxaborole treatment to food packaging material further comprisesembedding the benzoxaborole compound into the food packaging material,impregnating the food packaging material with the benzoxaborolecompound, or coating the food packaging material with the benzoxaborolecompound.
 10. The method of claim 6, wherein the food packaging materialis a chamber.
 11. The method of claim 5, wherein the chamber is aclamshell.
 12. The method of claim 11, wherein the clamshell comprisespolyethylene terephthalate.
 13. The method of claim 5, wherein thechamber further comprises a liquid-absorbing material.
 14. The method ofclaim 13, wherein the liquid-absorbing material is comprised in achamber component selected from the group consisting of a liner, awrapping, a label, a tag, a sticker, and a pad.
 15. The method of claim13, wherein the liquid-absorbing material provides for quick-release orslow-release of the benzoxaborole treatment to the food productcomprised in the chamber over a time period.
 16. The method of claim 10,wherein the chamber is a clamshell.
 17. The method of claim 16, whereinthe clamshell comprises polyethylene terephthalate.
 18. The method ofclaim 10, wherein the chamber further comprises a liquid-absorbingmaterial.
 19. The method of claim 18, wherein the liquid-absorbingmaterial is comprised in a chamber component selected from the groupconsisting of a liner, a wrapping, a label, a tag, a sticker, and a pad.20. The method of claim 18, wherein the liquid-absorbing materialprovides for quick-release or slow-release of the benzoxaboroletreatment to the food product comprised in the chamber over a timeperiod.